Screening for human papillomavirus (HPV) integration into host cell chromosomes typically requires large amounts of time and reagents. We developed a rapid and sensitive assay based on exonuclease V ...(ExoV) and quantitative polymerase chain reaction (qPCR) to determine HPV genome configurations in cell lines and tissues. We established the assay using genomic DNA from cell lines known to harbor integrated or episomal HPV16. DNA was incubated with ExoV, which is specific for linear DNA, and the DNA fraction resistant to digestion was measured by qPCR. The percent of DNA resistant to ExoV digestion was calculated relative to undigested DNA for determination of episomal or integrated HPV16. The ExoV assay was accurate, capable of distinguishing episomal from integrated HPV16 in cell lines and tissues. Future applications of the ExoV assay may include screening of HPV genome configurations in the progression of HPV-associated cancers.
•The ExoV assay is rapid and sensitive in detecting episomal and integrated HPV DNA.•The ExoV assay with HPV copy number identified episomal from type 1 or 2 integrants.•The ExoV assay monitors the physical state of HPV16 in organotypic raft tissues.
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GEOZS, IJS, IMTLJ, KILJ, KISLJ, NLZOH, NUK, OILJ, PNG, SAZU, SBCE, SBJE, UILJ, UL, UM, UPCLJ, UPUK, ZAGLJ, ZRSKP
The angiogenic capacity of local tissue critically regulates the response to ischemic injury. Elevated reactive oxygen species production, commonly associated with ischemic injury, has been shown to ...promote phosphorylation of the vascular endothelial growth factor receptor 2 (VEGFR2), a critical regulator of angiogenesis. Previous data from our lab demonstrated that diminished levels of the antioxidant glutathione positively augment ischemic angiogenesis. Here, we sought to determine the relationship between glutathione levels and oxidative stress in VEGFR2 signaling. We reveal that decreasing the ratio of GSH to GSSG with diamide leads to enhanced protein S-glutathionylation, increased reactive oxygen species (ROS) production, and enhanced VEGFR2 activation. However, increasing ROS alone was insufficient in activating VEGFR2, while ROS enhanced VEGF-stimulated VEGFR2 activation at supraphysiological levels. We also found that inhibiting glutathione reductase activity is sufficient to increase VEGFR2 activation and sensitizes cells to ROS-dependent VEGFR2 activation. Taken together, these data suggest that regulation of the cellular GSH:GSSG ratio critically regulates VEGFR2 activation. This work represents an important first step in separating thiol mediated signaling events from ROS dependent signaling.
Changes in GSH:GSSG but not ROS alone activates VEGFR2. This is a brief summary of the manuscript demonstrating that the addition of exogenous H2O2 is insufficient to activate VEGFR2; however decreasing GSH:GSSG results in ligand independent activation of VEGFR2. Display omitted
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GEOZS, IJS, IMTLJ, KILJ, KISLJ, NLZOH, NUK, OILJ, PNG, SAZU, SBCE, SBJE, UILJ, UL, UM, UPCLJ, UPUK, ZAGLJ, ZRSKP
Abstract
The mechanical loss of amorphous thin films of germania (GeO
2
) and titania-doped germania (Ti:GeO
2
) deposited by ion-beam sputtering onto silicon double-paddle oscillators was studied ...from 10 K to 290 K. Undoped germania was found to show a wide cryogenic mechanical loss peak centered at
∼
60
K with
ϕ
=
3.1
×
10
−
3
, which decreases to
1.1
×
10
−
3
as Ti-concentration increases to 44%. In addition to decreasing the height of this low-temperature peak, Ti-doping increases its width, and shifts its position toward lower temperatures. Annealing reduces the room temperature mechanical loss of Ti:GeO
2
and increases its cryogenic mechanical loss, which is consistent with trends observed in most amorphous oxides.
Understanding the local atomic order in amorphous thin film coatings and how it relates to macroscopic performance factors, such as mechanical loss, provides an important path towards enabling the ...accelerated discovery and development of improved coatings. High precision x-ray scattering measurements of thin films of amorphous zirconia-doped tantala (ZrO_{2}-Ta_{2}O_{5}) show systematic changes in intermediate range order (IRO) as a function of postdeposition heat treatment (annealing). Atomic modeling captures and explains these changes, and shows that the material has building blocks of metal-centered polyhedra and the effect of annealing is to alter the connections between the polyhedra. The observed changes in IRO are associated with a shift in the ratio of corner-sharing to edge-sharing polyhedra. These changes correlate with changes in mechanical loss upon annealing, and suggest that the mechanical loss can be reduced by developing a material with a designed ratio of corner-sharing to edge-sharing polyhedra.
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CMK, CTK, FMFMET, IJS, NUK, PNG, UL, UM
Brownian thermal noise as a result of mechanical loss in optical coatings will become the dominant source of noise at the most sensitive frequencies of ground-based gravitational-wave detectors. ...Experiments found, however, that a candidate material, amorphous Ta2O5, is unable to form an ultrastable glass and, consequently, to yield a film with significantly reduced mechanical loss through elevated-temperature deposition alone. X-ray scattering PDF measurements are carried out on films deposited and subsequently annealed at various temperatures. Inverse atomic modeling is used to analyze the short and medium range features in the atomic structure of these films. Furthermore, in silico deposition simulations of Ta2O5 are carried out at various substrate temperatures and an atomic level analysis of the growth at high temperatures is presented. It is observed that upon elevated-temperature deposition, short range features remain identical, whereas medium range order increases. After annealing, however, both the short and medium range orders of films deposited at different substrate temperatures are nearly identical. A discussion on the surface diffusion and glass transition temperatures indicates that future pursuits of ultrastable low-mechanical-loss films through elevated temperature deposition should focus on materials with a high surface mobility, and/or lower glass transition temperatures in the range of achievable deposition temperatueres.
ABSTRACT DXL (Diffuse X-ray emission from the Local Galaxy) is a sounding rocket mission designed to estimate the contribution of solar wind charge eXchange (SWCX) to the diffuse X-ray background and ...to help determine the properties of the Local Hot Bubble. The detectors are large area thin-window proportional counters with a spectral response that is similar to that of the PSPC used in the ROSAT All Sky Survey (RASS). A direct comparison of DXL and RASS data for the same part of the sky viewed from quite different vantage points in the solar system, and the assumption of approximate isotropy for the solar wind, allowed us to quantify the SWCX contribution to all six RASS bands (R1-R7, excluding R3). We find that the SWCX contribution at , where the DXL path crosses the Galactic plane, is for R1, for R2, for R4, for R5, and negligible for the R6 and R7 bands. Reliable models for the distribution of neutral H and He in the solar system permit estimation of the contribution of interplanetary SWCX emission over the the whole sky and correction of the RASS maps. We find that the average SWCX contribution in the whole sky is for R1, for R2, for R4, for R5, and negligible for R6 and R7.
Abstract
Introduction/Objective
CDC recommends the use of 4th generation HIV test as the first line of screening. Alere Determine HIV – 1/2 Ag/Ab Combo (Abbott Diagnostics Scarborough, Inc., ...Scarborough, Maine) (Alere) is a qualitative 4thgeneration assay that detects HIV 1/2 antibodies and p24 antigen. In this validation study, Alere was compared with OraQuick Advance Rapid HIV 1/2 Antibody test (OraSure Technologies, Inc., Bethlehem, PA) (Oraquick), a qualitative 3rd generation test and BioPlex (Bio-Rad Laboratories, Hercules, California) a 5th generation test that detects and differentiates between HIV 1 and 2 antibodies and p24 antigen.
Methods/Case Report
Comparison study: Forty-five samples were tested simultaneously on the three assays. Results for the patient samples were confirmed with Geenius (Bio- Rad Marnes-La-Coquette, France) or PCR assays for antibody or antigen positive cases respectively.
Precision study
A HIV-1 antibody positive and an all-negative sample were tested 10 times each to validate the precision of the Alere test.
Dilution study
A positive sample for HIV-1 antibody was used for dilution with a HIV negative serum sample. Diluted samples from 1:10 to 1:1000 were tested using Alere and BioPlex.
Results (if a Case Study enter NA)
Comparison study revealed that sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) for Alere test were 100%. Sensitivity for Oraquick was 86.6% and NPV was 77.77%. Specificity and PPV were 100%. BioPlex showed sensitivity and NPV of 100%, but specificity was 78.5% and PPV was 90.90%. Precision study showed reproducible results on the Alere test. Dilution study revealed that Alere assay gave positive results up to 1:300 dilution, and BioPlex assay was positive up to 1:500 dilution.
Conclusion
Our studies showed that Alere had the sensitivity and specificity of 100% making it an effective screening test in acute settings. BioPlex assay demonstrated that it was the most sensitive test with quantitative index for the HIV 1 and 2 antibodies and p24 antigen. Therefore, both Alere and BioPlex tests are good initial screening tests
Molecular oxygen (O2) is an essential component for survival and development. Variation in O2 levels leads to changes in molecular signaling and ultimately affects the physiological functions of many ...organisms. Nitric oxide (NO) and hydrogen sulfide (H2S) are two gaseous cellular signaling molecules that play key roles in several physiological functions involved in maintaining vascular homeostasis including vasodilation, anti-inflammation, and vascular growth. Apart from the aforementioned functions, NO and H2S are believed to mediate hypoxic responses and serve as O2 chemosensors in biological systems. In this literature review, we briefly discuss NO and H2S and their roles during hypoxia.
The effectiveness of a recently invented "steam pasteurization" (S) process in reducing pathogenic bacterial populations on surfaces of freshly slaughtered beef was determined and compared with that ...of other standard commercial methods including knife trimming (T), water washing (35 degrees C; W), hot water/steam vacuum spot cleaning (V), and spraying with 2% vol/vol lactic acid (54 degrees C, pH 2.25; L). These decontamination treatments were tested individually and in combinations. Cutaneus trunci muscles from freshly slaughtered steers were inoculated with feces containing Listeria monocytogenes Scott A, Escherichia coli O157:H7, and Salmonella typhimurium over a predesignated meat surface area, resulting in initial populations of ca. 5 log CFU/cm2 of each pathogen. Tissue samples were excised from each portion before and after decontamination treatments, and mean population reductions were determined. Treatment combinations evaluated were the following (treatment designations within the abbreviations indicate the order of application): TW, TWS, WS, VW, VWS, TWLS, and VWLS. These combinations resulted in reductions ranging from 3.5 to 5.3 log CFU/cm2 in all three pathogen populations. The TW, TWS, WS, TWLS, and VWLS combinations were equally effective (P 0.05), resulting in reductions ranging from 4.2 to 5.3 log CFU/cm2. When used individually, T, V, and S resulted in pathogen reductions ranging from 2.5 to 3.7 log CFU/cm2. Steam pasteurization consistently provided numerically greater pathogen reductions than T or V. Treatments T, V, and S were all more effective than W (which gave a reduction on the order of 1.0 log CFU/cm2). Steam pasteurization is an effective method for reducing pathogenic bacterial populations on surfaces of freshly slaughtered beef, with multiple decontamination procedures providing greatest overall reductions
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CEKLJ, GEOZS, IJS, IMTLJ, KILJ, KISLJ, NLZOH, NUK, OILJ, PNG, SAZU, SBCE, SBJE, UILJ, UL, UM, UPCLJ, UPUK, ZAGLJ, ZRSKP
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