The human papillomavirus type 16 E7 oncoprotein targets Myc-interacting zinc-finger protein-1

AbstractWe demonstrate that HPV-16 E7 forms a complex with Miz-1. UV-induced expression of the CDK-inhibitor p21Cip1 and subsequent cell cycle arrest depends upon endogenous Miz-1 in HPV-negative C33A cervical cancer cells containing mutated p53. Transient expression of E7 in C33A inhibits UV-induced expression of p21Cip1 and overcomes Miz-1-induced G1-phase arrest. The C-terminal E7Δ79LEDLL83-mutant with reduced Miz-1-binding capacity was impaired in its capability to repress p21Cip1 expression; whereas the pRB-binding-deficient E7C24G-mutant inhibited p21Cip1 expression similar to wild-type E7. Using ChIP, we demonstrate that endogenous E7 is bound to the endogenous p21Cip1 core-promoter in CaSki cells and RNAi-mediated knock down of Miz-1 abrogates E7-binding to the p21Cip1 promoter. Co-expression of E7 with Miz-1 inhibited Miz-1-induced p21Cip1 expression from the minimal-promoter via Miz-1 DNA-binding sites. Co-expression of E7Δ79LEDLL83 did not inhibit Miz-1-induced p21Cip1 expression. E7C24G retained E7-wild-type capability to inhibit Miz-1-dependent transactivation. These findings suggest that HPV-16 E7 can repress Miz-1-induced p21Cip1 gene expression

Characterization of DLK1(PREF1)+/CD34+ cells in vascular stroma of human white adipose tissue

AbstractSorting of native (unpermeabilized) SVF-cells from human subcutaneous (s)WAT for cell surface staining (cs) of DLK1 and CD34 identified three main populations: ~10% stained cs-DLK1+/cs-CD34−, ~20% cs-DLK1+/cs-CD34+dim and ~45% cs-DLK1−/cs-CD34+. FACS analysis after permeabilization showed that all these cells stained positive for intracellular DLK1, while CD34 was undetectable in cs-DLK1+/cs-CD34− cells. Permeabilized cs-DLK1−/cs-CD34+ cells were positive for the pericyte marker α-SMA and the mesenchymal markers CD90 and CD105, albeit CD105 staining was dim (cs-DLK1−/cs-CD34+/CD90+/CD105+dim/α-SMA+/CD45−/CD31−). Only these cells showed proliferative and adipogenic capacity. Cs-DLK1+/cs-CD34− and cs-DLK1+/cs-CD34+dim cells were also α-SMA+ but expressed CD31, had a mixed hematopoietic and mesenchymal phenotype, and could neither proliferate nor differentiate into adipocytes. Histological analysis of sWAT detected DLK1+/CD34+ and DLK1+/CD90+ cells mainly in the outer ring of vessel-associated stroma and at capillaries. DLK1+/α-SMA+ cells were localized in the CD34− perivascular ring and in adventitial vascular stroma. All these DLK1+ cells possess a spindle-shaped morphology with extremely long processes. DLK1+/CD34+ cells were also detected in vessel endothelium. Additionally, we show that sWAT contains significantly more DLK1+ cells than visceral (v)WAT. We conclude that sWAT has more DKL1+ cells than vWAT and contains different DLK1/CD34 populations, and only cs-DLK1−/cs-CD34+/CD90+/CD105+dim/α-SMA+/CD45−/CD31− cells in the adventitial vascular stroma exhibit proliferative and adipogenic capacity

Hydrology in the Sea of Marmara during the last 23 ka : implications for timing of Black Sea connections and sapropel deposition

Author Posting. © American Geophysical Union, 2010. This article is posted here by permission of American Geophysical Union for personal use, not for redistribution. The definitive version was published in Paleoceanography 25 (2010): PA1205, doi:10.1029/2009PA001735.Sediments deposited under lacustrine and marine conditions in the Sea of Marmara hold a Late Quaternary record for water exchange between the Black Sea and the Mediterranean Sea. Here we report a multiproxy data set based on oxygen and strontium isotope results obtained from carbonate shells, major and trace elements, and specific organic biomarker measurements, as well as a micropaleontological study from a 14C-dated sediment core retrieved from the Sea of Marmara. Pronounced changes occurred in δ18O and 87Sr/86Sr values at the fresh and marine water transition, providing additional information in relation to micropaleontological data. Organic biomarker concentrations documented the marine origin of the sapropelic layer while changes in n-alkane concentrations clearly indicated an enhanced contribution for organic matter of terrestrial origin before and after the event. When compared with the Black Sea record, the results suggest that the Black Sea was outflowing to the Sea of Marmara from the Last Glacial Maximum until the warmer Bølling-Allerød. The first marine incursion in the Sea of Marmara occurred at 14.7 cal ka B.P. However, salinification of the basin was gradual, indicating that Black Sea freshwaters were still contributing to the Marmara seawater budget. After the Younger Dryas (which is associated with a high input of organic matter of terrestrial origin) both basins were disconnected, resulting in a salinity increase in the Sea of Marmara. The deposition of organic-rich sapropel that followed was mainly related to enhanced primary productivity characterized by a reorganization of the phytoplankton population.We acknowledge support from INSU and the French Polar Institute IPEV

Precision of the current methods to measure the alkenone proxy UK'37 and absolute alkenone abundance in sediments : results of an interlaboratory comparison study

Measurements of the UK'37 index and the absolute abundance of alkenones in marine sediments are increasingly used in paleoceanographic research as proxies of past sea surface temperature and haptophyte (mainly coccolith-bearing species) primary productivity, respectively. An important aspect of these studies is to be able to compare reliably data obtained by different laboratories from a wide variety of locations. Hence the intercomparability of data produced by the research community is essential. Here we report results from an anonymous interlaboratory comparison study involving 24 of the leading laboratories that carry out alkenone measurements worldwide. The majority of laboratories produce data that are intercomparable within the considered confidence limits. For the measurement of alkenone concentrations, however, there are systematic biases between laboratories, which might be related to the techniques employed to quantify the components. The maximum difference between any two laboratories for any two single measurements of UK'37 in sediments is estimated, with a probability of 95%, to be <2.18C. In addition, the overall within-laboratory precision for the UK'37 temperature estimates is estimated to be <1.68C (95% probability). Similarly, from the analyses of alkenone concentrations the interlaboratory reproducibility is estimated at 32%, and the repeatability is estimated at 24%. The former is compared to a theoretical estimate of reproducibility and found to be excessively high. Hence there is certainly scope and a demonstrable need to improve reproducibility and repeatability of UK'37 and especially alkenone quantification data across the community of scientists involved in alkenone research

Precision of the current methods to measure the alkenone proxy U₃₇ K’ and absolute alkenone abundance in sediments: results of an interlaboratory comparison study

Measurements of the U₃₇ K’ index and the absolute abundance of alkenones in marine sediments are increasingly used in paleoceanographic research as proxies of past sea surface temperature and haptophyte (mainly coccolith-bearing species) primary productivity, respectively. An important aspect of these studies is to be able to compare reliably data obtained by different laboratories from a wide variety of locations. Hence the intercomparability of data produced by the research community is essential. Here we report results from an anonymous interlaboratory comparison study involving 24 of the leading laboratories that carry out alkenone measurements worldwide. The majority of laboratories produce data that are intercomparable within the considered confidence limits. For the measurement of alkenone concentrations, however, there are systematic biases between laboratories, which might be related to the techniques employed to quantify the components. The maximum difference between any two laboratories for any two single measurements of U₃₇ K’ in sediments is estimated, with a probability of 95%, to be <2.1°C. In addition, the overall within-laboratory precision for the U₃₇ K’ temperature estimates is estimated to be <1.6°C (95% probability). Similarly, from the analyses of alkenone concentrations the interlaboratory reproducibility is estimated at 32%, and the repeatability is estimated at 24%. The former is compared to a theoretical estimate of reproducibility and found to be excessively high. Hence there is certainly scope and a demonstrable need to improve reproducibility and repeatability of U₃₇ K’ and especially alkenone quantification data across the community of scientists involved in alkenone research