Identification of Domains of the HPV11 E1 Protein Required for DNA Replication in Vitro

AbstractThe HPV E1 and E2 proteins along with cellular factors, are required for replication of the viral genome. In this study we show that in vitro synthesized HPV11 E1 can support DNA replication in a cell-free system and is able to cooperate with E2 to recruit the host polymerase α primase to the HPV origin in vitro. Deletion analysis revealed that the N-terminal 166 amino acids of E1, which encompass a nuclear localization signal and a cyclin E-binding motif, are dispensable for E1-dependent DNA replication and for recruitment of pol α primase to the origin in vitro. A shorter E1 protein lacking the N-terminal 190 amino acids supported cell-free DNA replication at less than 25% the efficiency of wild-type E1 and was active in the pol α primase recruitment assay. An even shorter E1 protein lacking a functional DNA-binding domain due to a truncation of the N-terminal 352 amino acids was inactive in both assays despite the fact that it retains the ability to associate with E2 or pol α primase in the absence of ori DNA. We provide additional functional evidence that E1 interacts with pol α primase through the p70 subunit of the complex by showing that p70 can be recruited to the HPV origin by E1 and E2 in vitro, that the domain of E1 (amino acids 353–649) that binds to pol α primase in vitro is the same as that needed for interaction with p70 in the yeast two-hybrid system, and that exogenously added p70 competes with the interaction between E1 and pol α primase and inhibits E1-dependent cell-free DNA replication. On the basis of these results and the observation that pol α primase competes with the interaction between E1 and E2 in solution, we propose that these three proteins assemble at the origin in a stepwise process during which E1, following its interaction with E2, must bind to DNA prior to interacting with pol α primase

Development of the Barriers to Error Disclosure Assessment Tool

OBJECTIVES: An interprofessional group of health colleges' faculty created and piloted the Barriers to Error Disclosure Assessment tool as an instrument to measure barriers to medical error disclosure among health care providers. METHODS: A review of the literature guided the creation of items describing influences on the decision to disclose a medical error. Local and national experts in error disclosure used a modified Delphi process to gain consensus on the items included in the pilot. After receiving university institutional review board approval, researchers distributed the tool to a convenience sample of physicians (n = 19), pharmacists (n = 20), and nurses (n = 20) from an academic medical center. Means and SDs were used to describe the sample. Intraclass correlation coefficients were used to examine test-retest correspondence between the continuous items on the scale. Factor analysis with varimax rotation was used to determine factor loadings and examine internal consistency reliability. Cronbach α coefficients were calculated during initial and subsequent administrations to assess test-retest reliability. RESULTS: After omitting 2 items with intraclass correlation coefficient of less than 0.40, intraclass correlation coefficients ranged from 0.43 to 0.70, indicating fair to good test-retest correspondence between the continuous items on the final draft. Factor analysis revealed the following factors during the initial administration: confidence and knowledge barriers, institutional barriers, psychological barriers, and financial concern barriers to medical error disclosure. α Coefficients of 0.85 to 0.93 at time 1 and 0.82 to 0.95 at time 2 supported test-retest reliability. CONCLUSIONS: The final version of the 31-item tool can be used to measure perceptions about abilities for disclosing, impressions regarding institutional policies and climate, and specific barriers that inhibit disclosure by health care providers. Preliminary evidence supports the tool's validity and reliability for measuring disclosure variables

The surface distributions of the production of the major volatile species, H₂O, CO₂, CO and O₂, from the nucleus of comet 67P/Churyumov-Gerasimenko throughout the Rosetta Mission as measured by the ROSINA double focusing mass spectrometer

The Rosetta Orbiter Spectrometer for Ion and Neutral Analysis (ROSINA) suite of instruments operated throughout the over two years of the Rosetta mission operations in the vicinity of comet 67P/Churyumov-Gerasimenko. It measured gas densities and composition throughout the comet's atmosphere, or coma. Here we present two-years' worth of measurements of the relative densities of the four major volatile species in the coma of the comet, H₂O, CO₂, CO and O₂, by one of the ROSINA sub-systems called the Double Focusing Mass Spectrometer (DFMS). The absolute total gas densities were provided by the Comet Pressure Sensor (COPS), another ROSINA sub-system. DFMS is a very high mass resolution and high sensitivity mass spectrometer able to resolve at a tiny fraction of an atomic mass unit. We have analyzed the combined DFMS and COPS measurements using an inversion scheme based on spherical harmonics that solves for the distribution of potential surface activity of each species as the comet rotates, changing solar illumination, over short time intervals and as the comet changes distance from the sun and orientation of its spin axis over long time intervals. We also use the surface boundary conditions derived from the inversion scheme to simulate the whole coma with our fully kinetic Direct Simulation Monte Carlo model and calculate the production rates of the four major species throughout the mission. We compare the derived production rates with revised remote sensing observations by the Visible and Infrared Thermal Imaging Spectrometer (VIRTIS) as well as with published observations from the Microwave Instrument for the Rosetta Orbiter (MIRO). Finally we use the variation of the surface production of the major species to calculate the total mass loss over the mission and, for different estimates of the dust/gas ratio, calculate the variation of surface loss all over the nucleus

Interacting fermions in self-similar potentials

We consider interacting spinless fermions in one dimension embedded in self-similar quasiperiodic potentials. We examine generalizations of the Fibonacci potential known as precious mean potentials. Using a bosonization technique and a renormalization group analysis, we study the low-energy physics of the system. We show that it undergoes a metal-insulator transition for any filling factor, with a critical interaction that strongly depends on the position of the Fermi level in the Fourier spectrum of the potential. For some positions of the Fermi level the metal-insulator transition occurs at the non interacting point. The repulsive side is an insulator with a gapped spectrum whereas in the attractive side the spectrum is gapless and the properties of the system are described by a Luttinger liquid. We compute the transport properties and give the characteristic exponents associated to the frequency and temperature dependence of the conductivity.Comment: 18 pages, 10 EPS figure

Gene expression profiling in sinonasal adenocarcinoma

<p>Abstract</p> <p>Background</p> <p>Sinonasal adenocarcinomas are uncommon tumors which develop in the ethmoid sinus after exposure to wood dust. Although the etiology of these tumors is well defined, very little is known about their molecular basis and no diagnostic tool exists for their early detection in high-risk workers.</p> <p>Methods</p> <p>To identify genes involved in this disease, we performed gene expression profiling using cancer-dedicated microarrays, on nine matched samples of sinonasal adenocarcinomas and non-tumor sinusal tissue. Microarray results were validated by quantitative RT-PCR and immunohistochemistry on two additional sets of tumors.</p> <p>Results</p> <p>Among the genes with significant differential expression we selected <it>LGALS4, ACS5, CLU, SRI and CCT5 </it>for further exploration. The overexpression of <it>LGALS4, ACS5, SRI</it>, <it>CCT5 </it>and the downregulation of <it>CLU </it>were confirmed by quantitative RT-PCR. Immunohistochemistry was performed for LGALS4 (Galectin 4), ACS5 (Acyl-CoA synthetase) and CLU (Clusterin) proteins: LGALS4 was highly up-regulated, particularly in the most differentiated tumors, while CLU was lost in all tumors. The expression of ACS5, was more heterogeneous and no correlation was observed with the tumor type.</p> <p>Conclusion</p> <p>Within our microarray study in sinonasal adenocarcinoma we identified two proteins, LGALS4 and CLU, that were significantly differentially expressed in tumors compared to normal tissue. A further evaluation on a new set of tissues, including precancerous stages and low grade tumors, is necessary to evaluate the possibility of using them as diagnostic markers.</p

South Atlantic intermediate water advances into the North-east Atlantic with reduced Atlantic meridional overturning circulation during the last glacial period

The Nd isotopic composition (epsilon Nd) of seawater and cold-water coral (CWC) samples from the Gulf of Cadiz and the Alboran Sea, at a depth of 280-827 m were investigated in order to constrain middepth water mass dynamics within the Gulf of Cadiz over the past 40 ka. epsilon Nd of glacial and Holocene CWC from the Alboran Sea and the northern Gulf of Cadiz reveals relatively constant values (-8.6 to -9.0 and -9.5 to -10.4, respectively). Such values are similar to those of the surrounding present-day middepth waters from the Mediterranean Outflow Water (MOW; epsilon Nd approximate to -9.4) and Mediterranean Sea Water (MSW; epsilon Nd approximate to -9.9). In contrast, glacial epsilon Nd values for CWC collected at thermocline depth (550-827 m) in the southern Gulf of Cadiz display a higher average value (-8.90.4) compared to the present-day value (-11.70.3). This implies a higher relative contribution of water masses of Mediterranean (MSW) or South Atlantic origin (East Antarctic Intermediate Water, EAAIW). Our study has produced the first evidence of significant radiogenic epsilon Nd values (approximate to -8) at 19, 23-24, and 27 ka, which are coeval with increasing iceberg discharges and a weakening of Atlantic Meridional Overturning Circulation (AMOC). Since MOW epsilon Nd values remained stable during the last glacial period, it is suggested that these radiogenic epsilon Nd values most likely reflect an enhanced northward propagation of glacial EAAIW into the eastern Atlantic Basin

Involvement of the Intrinsic/Default System in Movement-Related Self Recognition

The question of how people recognize themselves and separate themselves from the environment and others has long intrigued philosophers and scientists. Recent findings have linked regions of the ‘default brain’ or ‘intrinsic system’ to self-related processing. We used a paradigm in which subjects had to rely on subtle sensory-motor synchronization differences to determine whether a viewed movement belonged to them or to another person, while stimuli and task demands associated with the “responded self” and “responded other” conditions were precisely matched. Self recognition was associated with enhanced brain activity in several ROIs of the intrinsic system, whereas no differences emerged within the extrinsic system. This self-related effect was found even in cases where the sensory-motor aspects were precisely matched. Control conditions ruled out task difficulty as the source of the differential self-related effects. The findings shed light on the neural systems underlying bodily self recognition

Gene expression profiling of rat spermatogonia and Sertoli cells reveals signaling pathways from stem cells to niche and testicular cancer cells to surrounding stroma

Background: Stem cells and their niches are studied in many systems, but mammalian germ stem cells (GSC) and their niches are still poorly understood. In rat testis, spermatogonia and undifferentiated Sertoli cells proliferate before puberty, but at puberty most spermatogonia enter spermatogenesis, and Sertoli cells differentiate to support this program. Thus, pre-pubertal spermatogonia might possess GSC potential and pre-pubertal Sertoli cells niche functions. We hypothesized that the different stem cell pools at pre-puberty and maturity provide a model for the identification of stem cell and niche-specific genes. We compared the transcript profiles of spermatogonia and Sertoli cells from pre-pubertal and pubertal rats and examined how these related to genes expressed in testicular cancers, which might originate from inappropriate communication between GSCs and Sertoli cells. Results: The pre-pubertal spermatogonia-specific gene set comprised known stem cell and spermatogonial stem cell (SSC) markers. Similarly, the pre-pubertal Sertoli cell-specific gene set comprised known niche gene transcripts. A large fraction of these specifically enriched transcripts encoded trans-membrane, extra-cellular, and secreted proteins highlighting stem cell to niche communication. Comparing selective gene sets established in this study with published gene expression data of testicular cancers and their stroma, we identified sets expressed genes shared between testicular tumors and pre-pubertal spermatogonia, and tumor stroma and pre-pubertal Sertoli cells with statistic significance. Conclusions: Our data suggest that SSC and their niche specifically express complementary factors for cell communication and that the same factors might be implicated in the communication between tumor cells and their micro-enviroment in testicular cancer

Associations of common breast cancer susceptibility alleles with risk of breast cancer subtypes in BRCA1 and BRCA2 mutation carriers

Introduction: More than 70 common alleles are known to be involved in breast cancer (BC) susceptibility, and several exhibit significant heterogeneity in their associations with different BC subtypes. Although there are differences in the association patterns between BRCA1 and BRCA2 mutation carriers and the general population for several loci, no study has comprehensively evaluated the associations of all known BC susceptibility alleles with risk of BC subtypes in BRCA1 and BRCA2 carriers. Methods: We used data from 15,252 BRCA1 and 8,211 BRCA2 carriers to analyze the associations between approximately 200,000 genetic variants on the iCOGS array and risk of BC subtypes defined by estrogen receptor (ER), progesterone receptor (PR), human epidermal growth factor receptor 2 (HER2) and triple-negative- (TN) status; morphologic subtypes; histological grade; and nodal involvement. Results: The estimated BC hazard ratios (HRs) for the 74 known BC alleles in BRCA1 carriers exhibited moderate correlations with the corresponding odds ratios from the general population. However, their associations with ER-positive BC in BRCA1 carriers were more consistent with the ER-positive as

Assessing associations between the AURKAHMMR-TPX2-TUBG1 functional module and breast cancer risk in BRCA1/2 mutation carriers

While interplay between BRCA1 and AURKA-RHAMM-TPX2-TUBG1 regulates mammary epithelial polarization, common genetic variation in HMMR (gene product RHAMM) may be associated with risk of breast cancer in BRCA1 mutation carriers. Following on these observations, we further assessed the link between the AURKA-HMMR-TPX2-TUBG1 functional module and risk of breast cancer in BRCA1 or BRCA2 mutation carriers. Forty-one single nucleotide polymorphisms (SNPs) were genotyped in 15,252 BRCA1 and 8,211 BRCA2 mutation carriers and subsequently analyzed using a retrospective likelihood appr