bluuebunny/crossref_metadata_2025_split huggingface dataset:
Role of the Specific Interaction of UL112-113 p84 with UL44 DNA Polymerase Processivity Factor in Promoting DNA Replication of Human Cytomegalovirus
Authors: Kim Young-Eui , Ahn Jin-Hyun 
Published: June 2010

ABSTRACT

The human cytomegalovirus (HCMV) UL112-113 region encodes four phosphoproteins with common amino termini (p34, p43, p50, and p84) via alternative splicing and is thought to be required for efficient viral DNA replication. We have previously shown that interactions among the four UL112-113 proteins regulate their intranuclear targeting and enable the recruitment of the UL44 DNA polymerase processivity factor to viral prereplication foci. Here, we show that in virus-infected cells, the UL112-113 proteins form a complex with UL44 and other replication proteins, such as UL84 and IE2.
In vitro
assays showed that all four phosphoproteins interacted with UL44. Interestingly, p84 required both the shared amino-terminal region and the specific near-carboxy-terminal region for UL44 binding. UL44 required both the carboxy-terminal region and the central region, including the dimerization domain for p84 binding. The production of recombinant virus from mutant Towne bacterial artificial chromosome (BAC) DNA, which encodes intact p34, p43, and p50 and a carboxy-terminally truncated p84 defective in UL44 binding, was severely impaired compared to wild-type BAC DNA. A similar defect was observed when mutant BAC DNA encoded a carboxy-terminally truncated UL44 defective in p84 binding. In cotransfection replication assays using six replication core proteins, UL84, IE2, and UL112-113, the efficient replication of an HCMV
ori
Lyt-containing plasmid required the regions of p84 and UL44 necessary for their interaction. Our data suggest that the UL112-113 proteins form a complex with other replication proteins such as UL44, UL84, and IE2 and that the specific interaction of UL112-113 p84 with UL44 is necessary for efficient viral DNA replication.
Differential Sensitivity of Well-Differentiated Avian Respiratory Epithelial Cells to Infection by Different Strains of Infectious Bronchitis Virus
Authors: Abd El Rahman Sahar , Winter Christine , El-Kenawy Ali , Neumann Ulrich , Herrler Georg 
Published: June 2010

ABSTRACT

Infectious bronchitis virus (IBV) is an avian coronavirus affecting the respiratory tract of chickens. To analyze IBV infection of the lower respiratory tract, we applied a technique that uses precision-cut lung slices (PCLSs). This method allows infection of bronchial cells within their natural tissue composition under
in vitro
conditions. We demonstrate that IBV strains 4/91, Italy02, and QX infect ciliated and mucus-producing cells of the bronchial epithelium, whereas cells of the parabronchial tissue are resistant to infection. This is the first study, using PCLSs of chicken origin, to analyze virus infection. PCLSs should also be a valuable tool for investigation of other respiratory pathogens, such as avian influenza viruses.
Immediate-Early Expression of a Recombinant Antigen by Modified Vaccinia Virus Ankara Breaks the Immunodominance of Strong Vector-Specific B8R Antigen in Acute and Memory CD8 T-Cell Responses
Authors: Baur Karen , Brinkmann Kay , Schweneker Marc , Pätzold Juliane , Meisinger-Henschel Christine , Hermann Judith , Steigerwald Robin , Chaplin Paul , Suter Mark , Hausmann Jürgen 
Published: June 2010

ABSTRACTEfficient T-cell responses against recombinant antigens expressed by vaccinia virus vectors require expression of these antigens in the early phase of the virus replication cycle. The kinetics of recombinant gene expression in poxviruses are largely determined by the promoter chosen. We used the highly attenuated modified vaccinia virus Ankara (MVA) to determine the role of promoters in the induction of CD8 T-cell responses. We constructed MVA recombinants expressing either enhanced green fluorescent protein (EGFP) or chicken ovalbumin (OVA), each under the control of a hybrid early-late promoter (pHyb) containing five copies of a strong early element or the well-known early-late p7.5 or pS promoter for comparison. In primary or cultured cells, EGFP expression under the control of pHyb was detected within 30 min, as an immediate-early protein, and remained higher over the first 6 h of infection than p7.5- or pS-driven EGFP expression. Repeated immunizations of mice with recombinant MVA expressing OVA under the control of the pHyb promoter led to superior acute and memory CD8 T-cell responses compared to those to p7.5- and pS-driven OVA. Moreover, OVA expressed under the control of pHyb replaced the MVA-derived B8R protein as the immunodominant CD8 T-cell antigen after three or more immunizations. This is the first demonstration of an immediate-early neoantigen expressed by a poxviral vector resulting in superior induction of neoantigen-specific CD8 T-cell responses.
Differential Effects of DNA Double-Strand Break Repair Pathways on Single-Strand and Self-Complementary Adeno-Associated Virus Vector Genomes
Authors: Cataldi Marcela P. , McCarty Douglas M. 
Published: June 2010

ABSTRACT

The linear DNA genomes of recombinant adeno-associated virus (rAAV) gene delivery vectors are acted upon by multiple DNA repair and recombination pathways upon release into the host nucleus, resulting in circularization, concatemer formation, or chromosomal integration. We have compared the fates of single-strand rAAV (ssAAV) and self-complementary AAV (scAAV) genomes in cell lines deficient in each of three signaling factors, ATM, ATR, and DNA-PK
CS
, orchestrating major DNA double-strand break (DSB) repair pathways. In cells deficient in ATM, transduction as scored by green fluorescent protein (GFP) expression is increased relative to that in wild-type (wt) cells by 2.6-fold for ssAAV and 6.6-fold for scAAV vectors, arguing against a mechanism related to second-strand synthesis. The augmented transduction is not reflected in Southern blots of nuclear vector DNA, suggesting that interactions with ATM lead to silencing in normal cells. The additional functional genomes in ATM
−/−
cells remain linear, and the number of circularized genomes is not affected by the mutation, consistent with compartmentalization of genomes into different DNA repair pathways. A similar effect is observed in ATR-deficient cells but is specific for ssAAV vector. Conversely, a large decrease in transduction is observed in cells deficient in DNA-PK
CS
, which is involved in DSB repair by nonhomologous end joining rather than homologous recombination. The mutations also have differential effects on chromosomal integration of ssAAV versus scAAV vector genomes. Integration of ssAAV was specifically reduced in ATM
−/−
cells, while scAAV integration was more profoundly inhibited in DNA-PK
CS
−/−
cells. Taken together, the results suggest that productive rAAV genome circularization is mediated primarily by nonhomologous end joining.
The Combination of a Histone Deacetylase Inhibitor with the Bcl-2 Homology Domain-3 Mimetic GX15-070 Has Synergistic Antileukemia Activity by Activating Both Apoptosis and Autophagy
Authors: Wei Yue , Kadia Tapan , Tong Weigang , Zhang Ming , Jia Yu , Yang Hui , Hu Yumin , Tambaro Francesco Paolo , Viallet Jean , O'Brien Susan , Garcia-Manero Guillermo 
Published: June 2010

AbstractPurpose: Single-agent histone deacetylase inhibitors (HDACi) have limited clinical activity in human leukemia. Although the way HDACi exert their antileukemia effect is not fully understood, it is accepted that induction of apoptosis is important. We hypothesized, therefore, that combination of an HDACi with a proapoptotic agent, such as the Bcl-2 homology domain-3 mimetic GX15-070, could result in enhanced antileukemia activity.Experimental Design: We analyzed the cellular and molecular effects of two different HDACi (MGCD0103 and vorinostat) in combination with GX15-070 in leukemia cell lines and primary acute myelogenous leukemia cells.Results: We showed that the combination had synergistic antileukemia effect both in leukemia cell lines and in primary acute myelogenous leukemia cells. Using molecular markers and electron microscopy, we observed that in addition to apoptosis, autophagy accounts for the nonapoptotic decrease in cell viability, an effect that could be inhibited by chloroquine, an inhibitor of autophagy. Finally, we established a role for calpain activity in the induction of both autophagy and apoptosis by this combination.Conclusions: The combination of an HDACi and GX15-070 has synergistic antileukemia activity, and the effect is mediated by induction of apoptosis and autophagy. The combination should be studied in clinical trials of leukemia and the role of autophagy in leukemia therapy needs to be better understood. Clin Cancer Res; 16(15); 3923–32. ©2010 AACR.
PIKing the Right Patient
Authors: Turke Alexa B. , Engelman Jeffrey A. 
Published: June 2010

Abstract
HER2 amplification and PIK3CA mutation were validated as biomarkers for sensitivity to the single-agent phosphoinositide 3-kinase (PI3K) inhibitor, GDC-0941, in breast cancer models. A novel expression profile was developed to identify other breast cancers sensitive to PI3K inhibitors. These expression studies highlighted feedback networks connecting TORC1, PI3K, and mitogen-activated protein kinase (MAPK) pathways, and underscored the potential for combination therapies. Clin Cancer Res; 16(14); 3523–5. ©2010 AACR.
Alterations of L-Type Calcium Current and Cardiac Function in CaMKIIδ Knockout Mice
Authors: Xu Lin , Lai Dongwu , Cheng Jun , Lim Hyun Joung , Keskanokwong Thitima , Backs Johannes , Olson Eric N. , Wang Yanggan 
Published: June 2010

Rationale:Recent studies have highlighted important roles of CaMKII in regulating Ca2+handling and excitation-contraction coupling. However, the cardiac effect of chronic CaMKII inhibition has not been well understood.Objective:We have tested the alterations of L-type calcium current (ICa) and cardiac function in CaMKIIδ knockout (KO) mouse left ventricle (LV).Methods and Results:We used the patch-clamp method to record ICain ventricular myocytes and found that in KO LV, basal ICawas significantly increased without changing the transmural gradient of ICadistribution. Substitution of Ba2+for Ca2+showed similar increase in IBa. There was no change in the voltage dependence of ICaactivation and inactivation. ICarecovery from inactivation, however, was significantly slowed. In KO LV, the Ca2+-dependent ICafacilitation (CDF) and ICaresponse to isoproterenol (ISO) were significantly reduced. However, ISO response was reversed by β2-adrenergic receptor (AR) inhibition. Western blots showed a decrease in β1-AR and an increase in Cav1.2, β2-AR, and Gαi3 protein levels. Ca2+transient and sarcomere shortening in KO myocytes were unchanged at 1-Hz but reduced at 3-Hz stimulation. Echocardiography in conscious mice revealed an increased basal contractility in KO mice. However, cardiac reserve to work load and β-adrenergic stimulation was reduced. Surprisingly, KO mice showed a reduced heart rate in response to work load or β-adrenergic stimulation.Conclusions:Our results implicate physiological CaMKII activity in maintaining normal ICa, Ca2+handling, excitation-contraction coupling, and the in vivo heart function in response to cardiac stress.
Comparison of Satellite Cloud Masks with Ceilometer Sky Conditions in Southern Finland
Authors: Joro Sauli , Hyvärinen Otto , Kotro Janne 
Published: June 2010

AbstractThe cloud mask is an essential product derived from satellite data. Whereas cloud analysis applications typically make use of information from cloudy pixels, many other applications require cloud-free conditions. For this reason many organizations have their own cloud masks tuned to serve their particular needs. Being a fundamental product, continuous quality monitoring and validation of these cloud masks are vital. This study evaluated the performance of the European Organisation for the Exploitation of Meteorological Satellites (EUMETSAT) Meteorological Products Extraction Facility cloud mask (MPEF), together with the Nowcasting Satellite Application Facility (SAFNWC) cloud masks provided by Météo-France (SAFNWC/MSG) and the Swedish Meteorological and Hydrological Institute (SAFNWC/PPS), in the high-latitude area of greater Helsinki in Finland. The first two used the Spinning Enhanced Visible and Infrared Imager (SEVIRI) instrument from the geostationary Meteosat-8 satellite, whereas the last used the Advanced Very High Resolution Radiometer (AVHRR) instrument on board the polar-orbiting NOAA satellite series. Ceilometer data from the Helsinki Testbed, an extensive observation network covering the greater Helsinki area in Finland, were used as reference data in the cloud mask comparison. A computational method, called bootstrapping, is introduced to account for the strong temporal and spatial correlation of the ceilometer observations. The method also allows the calculation of the confidence intervals (CI) for the results. This study comprised data from February and August 2006. In general, the SAFNWC/MSG algorithm performed better than MPEF. Differences were found especially in the early morning low cloud detection. The SAFNWC/PPS cloud mask performed very well in August, better than geostationary-based masks, but had problems in February when its performance was worse. The use of the CIs gave the results more depth, and their use should be encouraged.
Mechanisms Affecting the Transition from Shallow to Deep Convection over Land: Inferences from Observations of the Diurnal Cycle Collected at the ARM Southern Great Plains Site
Authors: Zhang Yunyan , Klein Stephen A. 
Published: June 2010

AbstractSummertime observations for 11 yr from the Atmospheric Radiation Measurement (ARM) Climate Research Facility Southern Great Plains (SGP) site are used to investigate mechanisms controlling the transition from shallow to deep convection over land. It is found that a more humid environment immediately above the boundary layer is present before the start of late afternoon heavy precipitation events. The higher moisture content is brought by wind from the south. Greater boundary layer inhomogeneity in moist static energy, temperature, moisture, and horizontal wind before precipitation begins is correlated to larger rain rates at the initial stage of precipitation. In an examination of afternoon rain statistics, higher relative humidity above the boundary layer is correlated to an earlier onset and longer duration of afternoon precipitation events, whereas greater boundary layer inhomogeneity and atmospheric instability in the 2–4-km layer above the surface are positively correlated to the total rain amount and the maximum rain rate. Although other interpretations may be possible, these observations are consistent with theories for the transition from shallow to deep convection that emphasize the role of a moist lower free troposphere and boundary layer inhomogeneity.
Mesoscale Cyclone Activity over the Ice-Free Southern Ocean: 1999–2008
Authors: Irving Damien , Simmonds Ian , Keay Kevin 
Published: June 2010

Abstract
Mesoscale cyclones play an important role in the weather and climate of the Southern Hemisphere (SH) mid-to-high latitudes. However, the relatively small size and short lifetime of these systems, combined with the lack of available conventional data in this region, means that there is a poor understanding of their climatological characteristics. In this study, the University of Melbourne cyclone-finding algorithm was applied to relatively high-resolution scatterometer-derived surface pressure fields, obtained from the Department of Atmospheric Sciences at the University of Washington, to produce a decade-long (1999–2008) seasonal climatology of mesoscale cyclone activity over the ice-free regions of the Southern Ocean.
The frequency of mesoscale cyclone activity was found to be highest just to the north of the sea ice zone, reaching a maximum over the Amundsen and Bellingshausen Seas (ABS), while the southern Indian Ocean was associated with systems of the largest depth, intensity, and momentum flux at the ocean surface. These spatial patterns in mean mesoscale cyclone characteristics showed a broad resemblance to those reported in existing synoptic-scale cyclone climatologies. Maximum wind speed data indicated that SH polar lows may be more frequent than the current literature suggests, while strong positive trends identified in mesoscale cyclone frequency over the ABS may represent a contributing factor to the rapid warming observed in that region over recent years. Partial correlation analyses indicated a link between mesoscale cyclone frequency and the southern annular mode.
Retracing the Path of the Sons of Hilal
Authors: Galley Micheline 
Published: June 2010

AbstractThe article aims to contribute to a wider knowledge of the Hilâl epic, a masterwork of popular Arabic literature that tells the story of a nomadic pastoral people from the Arabian deserts. The focus is on the ‘Taghrîba’ cycle, which relates the migration in the 11th century of these Sons of Hilâl to Ifrîqiyya, present-day Tunisia.In this context reference is made to the political act of the Fatimid power that launched the Hilalians on the conquest of Ifrîqiyya, as well as to historians’ and linguists’ perception of it. But interest is centred more on the texts, which are copiously quoted, their poetic quality and their message – the expression of a tribal humanism, composed of grandeur and human dignity, particular to those desert people.
Acquired mutations in the genes encoding IDH1 and IDH2 both are recurrent aberrations in acute myeloid leukemia: prevalence and prognostic value
Authors: Abbas Saman , Lugthart Sanne , Kavelaars François G. , Schelen Anita , Koenders Jasper E. , Zeilemaker Annelieke , van Putten Wim J. L. , Rijneveld Anita W. , Löwenberg Bob , Valk Peter J. M. 
Published: June 2010

Abstract
Somatic mutations in isocitrate dehydrogenase 1 and 2 (IDH1 and IDH2) were recently demonstrated in acute myeloid leukemia (AML), but their prevalence and prognostic impact remain to be explored in large extensively characterized AML series, and also in various other hematologic malignancies. Here, we demonstrate in 893 newly diagnosed cases of AML mutations in the IDH1 (6%) and IDH2 (11%) genes. Moreover, we identified IDH mutations in 2 JAK2 V617F myeloproliferative neoplasias (n = 96), a single case of acute lymphoblastic leukemia (n = 96), and none in chronic myeloid leukemias (n = 81). In AML, IDH1 and IDH2 mutations are more common among AML with normal karyotype and NPM1mutant genotypes. IDH1 mutation status is an unfavorable prognostic factor as regards survival in a composite genotypic subset lacking FLT3ITD and NPM1mutant. Thus, IDH1 and IDH2 mutations are common genetic aberrations in AML, and IDH1 mutations may carry prognostic value in distinct subtypes of AML.
Effect of vascular endothelial growth factor and its receptor KDR on the transendothelial migration and local trafficking of human T cells in vitro and in vivo
Authors: Edelbauer Monika , Datta Dipak , Vos Ingrid H. C. , Basu Aninda , Stack Maria P. , Reinders Marlies E. J. , Sho Masayuki , Calzadilla Katiana , Ganz Peter , Briscoe David M. 
Published: June 2010

AbstractIn these studies, we find that the vascular endothelial growth factor (VEGF) receptor KDR is expressed on subsets of mitogen-activated CD4+ and CD8+ T cells in vitro. We also found that KDR colocalizes with CD3 on mitogen-activated T cells in vitro and on infiltrates within rejecting human allografts in vivo. To evaluate whether VEGF and KDR mediate lymphocyte migration across endothelial cells (ECs), we used an in vitro live-time transmigration model and observed that both anti-VEGF and anti-KDR antibodies inhibit the transmigration of both CD4+ and CD8+ T cells across tumor necrosis factorα (TNFα)–activated, but not unactivated ECs. In addition, we found that interactions among CD4+ or CD8+ T cells and TNFα–activated ECs result in the induction of KDR on each T cell subset, and that KDR-expressing lymphocytes preferentially transmigrate across TNFα–activated ECs. Finally, using a humanized severe combined immunodeficient mouse model of lymphocyte trafficking, we found that KDR-expressing lymphocytes migrate into human skin in vivo, and that migration is reduced in mice treated with a blocking anti-VEGF antibody. These observations demonstrate that induced expression of KDR on subsets of T cells, and locally expressed VEGF, facilitate EC-dependent lymphocyte chemotaxis, and thus, the localization of T cells at sites of inflammation.
Notch1 as a potential therapeutic target in cutaneous T-cell lymphoma
Authors: Kamstrup Maria R. , Gjerdrum Lise Mette Rahbek , Biskup Edyta , Thyssing Lauenborg Britt , Ralfkiaer Elisabeth , Woetmann Anders , Ødum Niels , Gniadecki Robert 
Published: June 2010

AbstractDeregulation of Notch signaling has been linked to the development of T-cell leukemias and several solid malignancies. Yet, it is unknown whether Notch signaling is involved in the pathogenesis of mycosis fungoides and Sézary syndrome, the most common subtypes of cutaneous T-cell lymphoma. By immunohistochemistry of 40 biopsies taken from skin lesions of mycosis fungoides and Sézary syndrome, we demonstrated prominent expression of Notch1 on tumor cells, especially in the more advanced stages. The γ-secretase inhibitor I blocked Notch signaling and potently induced apoptosis in cell lines derived from mycosis fungoides (MyLa) and Sézary syndrome (SeAx, HuT-78) and in primary leukemic Sézary cells. Specific down-regulation of Notch1 (but not Notch2 and Notch3) by siRNA induced apoptosis in SeAx. The mechanism of apoptosis involved the inhibition of nuclear factor-κB, which is the most important prosurvival pathway in cutaneous T-cell lymphoma. Our data show that Notch is present in cutaneous T-cell lymphoma and that its inhibition may provide a new way to treat cutaneous T-cell lymphoma.
Genomic deletion of the whole IgH 3′ regulatory region (hs3a, hs1,2, hs3b, and hs4) dramatically affects class switch recombination and Ig secretion to all isotypes
Authors: Vincent-Fabert Christelle , Fiancette Remi , Pinaud Eric , Truffinet Véronique , Cogné Nadine , Cogné Michel , Denizot Yves 
Published: June 2010

Abstract
The immunoglobulin heavy chain locus (IgH) undergoes multiple changes along B-cell differentiation. In progenitor B cells, V(D)J assembly allows expression of μ heavy chains. In mature B cells, class switch recombination may replace the expressed constant (C)μ gene with a downstream CH gene. Finally, plasma cell differentiation strongly boosts IgH transcription. How the multiple IgH transcriptional enhancers tune these changes is unclear. Here we demonstrate that deletion of the whole IgH 3′ regulatory region (3′RR) allows normal maturation until the stage of IgM/IgD expressing lymphocytes, but nearly abrogates class switch recombination to all CH genes. Although plasma cell numbers are unaffected, we reveal the role of the 3′RR into the transcriptional burst normally associated with plasma cell differentiation. Our study shows that transcriptional changes and recombinations occurring after antigen-encounter appear mainly controlled by the 3′RR working as a single functional unit.