A rapid non-radioactive technique for measurement of repair synthesis in primary human fibroblasts by incorporation of ethynyl deoxyuridine (EdU)

Xeroderma pigmentosum (XP) is an autosomal recessive genetic disorder. Afflicted patients show extreme sun-sensitivity and skin cancer predisposition. XP is in most cases associated with deficient nucleotide excision repair (NER), which is the process responsible for removing photolesions from DNA. Measuring NER activity by nucleotide incorporation into repair patches, termed ‘unscheduled DNA synthesis (UDS)’, is one of the most commonly used assays for XP-diagnosis and NER research. We have established a rapid and accurate procedure for measuring UDS by replacement of thymidine with 5-ethynyl-2'-deoxyuridine (EdU). EdU incorporated into repair patches can be directly conjugated to fluorescent azide derivatives, thereby obviating the need for either radiolabeled thymidine or denaturation and antibody detection of incorporated bromodeoxyuridine (BrdU). We demonstrate that the EdU incorporation assay is compatible with conventional techniques such as immunofluorescent staining and labeling of cells with micro-latex beads. Importantly, we can complete the entire UDS assay within half a day from preparation of the assay coverslips; this technique may prove useful as a method for XP diagnosis

Ultraviolet radiation exposure to the face in patients with xeroderma pigmentosum and healthy controls:applying a novel methodology to define photoprotection behaviour*

BACKGROUND: In xeroderma pigmentosum (XP), the main means of preventing skin and eye cancers is extreme protection against ultraviolet radiation (UVR). Protection is most important for the face. OBJECTIVES: We aimed to assess how well patients with XP adhere to medical advice to protect against UVR by objectively estimating the mean daily dose of UVR to the face. METHODS: We objectively estimated the UVR dose to the face in 36 patients with XP and 25 healthy individuals over 3 weeks in the summer. We used a new methodology which combined UVR dose measurements from a wrist‐worn dosimeter with an activity diary record of face photoprotection behaviour for each 15‐min period spent outside. A protection factor was associated with each behaviour, and the data were analysed using a negative binomial mixed‐effects model. RESULTS: The mean daily UVR dose (weighted for DNA damage capacity) to the face in the patients with XP was 0·13 standard erythemal doses (SEDs) (mean in healthy individuals = 0·51 SED). There was wide variation between patients (range < 0·01–0·48 SED/day). Self‐caring adult patients had a very similar UVR dose to the face as cared‐for patients (0·13 vs. 0·12 SED/day), despite photoprotecting much more poorly when outside, because the self‐caring adults were outside in daylight much less. CONCLUSIONS: Photoprotection behaviour varies widely within the XP group indicating that nonadherence to photoprotection advice is a significant issue. The timing and duration of going outside are as important as photoprotective measures taken when outside, to determine the UVR exposure to the face. This new methodology will be of value in identifying the sources of UVR exposure in other conditions in which facial UVR exposure is a key outcome, particularly in patients with multiple nonmelanoma skin cancers

Aftermarketfailure: Windows XP\u27s End of Support

After 12 years, support for Windows XP will end on April 8, 2014. So proclaims a Microsoft website with a helpful clock counting down the days. What does this mean? the website asks. It means you should take action. You should migrate to a current supported operating system - such as Windows 8.1 - so you can receive regular security updates to protect [your] computer from malicious attacks. The costs of mass migration will be immense. About 30% of all desktop PCs are running Windows XP right now. An estimated 10% of the U.S. government\u27s computers run Windows XP, including thousands of computers on classified military and diplomatic networks. And the costs of staying put? They will be enormous, too. It turns out that 95% of the world\u27s ATMs are powered by Windows XP, and there is no readily available substitute in the offing. In one example of these exorbitant costs, the price of either extending support or upgrading to another version of Windows for each of Britain\u27s major banks will be in the hundreds of millions. Costs will be similar, or perhaps even higher, worldwide. The failure to continue to patch unpatched computers will also have serious ramifications for society. Hackers will keep scouring Windows XP for flaws, holes, and vulnerabilities for many years after Microsoft ends support for its vaunted operating system. Zero-day vulnerabilities-flaws hackers have long known about but have waited to exploit for fear that the vulnerability will be immediately patched (and could therefore only be used once or for a short time)-will now be exploitable in perpetuity. Experts have repeatedly warned that April 8 could spark a hacker feeding frenzy. Yet, ironically, those who run Windows XP pose a greater threat to others than to themselves. In the special case of governments, enterprise businesses, and financial institutions, failure to patch or migrate will expose the personal data of millions of individuals to theft, fraud, and abuse. When the wave of security breaches from unpatched machines arrives, the lawsuits against companies for failure to secure user data properly will become even more costly. There may also be immense blows to consumer confidence, leading to welfare-reducing market-wide substitutions away from e-commerce

Analyzing the Applicability of an Agile Methodology to Distributed Collaborative Software Development

Today, information technology (IT) has penetrated most domains of business and private life. The knitting of IT-systems and their dependencies are getting more complex every day. For businesses, this development can mean great opportunities. IT has become a main driver for competitive advantage and business success. On the other hand, misled software development (SD) projects can mean an existential threat to the operational and financial situation of a company. The efficient development of effective software is an essential part of optimally facing present and future challenges. Managing SD with traditional methodologies often leads to high planning and management overhead and still, severe schedule deviations and budget overruns cannot be eliminated. The sequential and plan-driven traditional approaches are often not able to support an adequate reaction to either internally or externally caused changes in requirements. Complex and unclear system landscapes with diverse interfaces, ambiguous customer requirements, changing business strategies or fluctuating legal requirements are just a few examples for possible sources of changing system requirements. Today, Extreme Programming (XP) is the most popular agile development methodology supported by the Agile Alliance. Its name was chosen because it claims to bring common sense to an extreme level. It focuses on communication, simplicity, feedback and courage, to improve the speed and quality of SD. Formal processes and documentation are neglected in favor of tacit knowledge to improve flexibility. Close communication between developers and the continuous integration of customer representatives are key components of XP. XP was initially developed for small to medium sized collocated development teams. This paper analyzes to what extent XP can be transferred to larger distributed developing endeavors. The focus is on XP, because it is the methodology with the highest congruence to the original Agile Manifesto. It does not claim to be all new, but to be an aligned composition of well established ideas and practices from other methodologies

Aftermarketfailure: Windows XP\u27s End of Support

After 12 years, support for Windows XP will end on April 8, 2014. So proclaims a Microsoft website with a helpful clock counting down the days. What does this mean? the website asks. It means you should take action. You should migrate to a current supported operating system - such as Windows 8.1 - so you can receive regular security updates to protect [your] computer from malicious attacks. The costs of mass migration will be immense. About 30% of all desktop PCs are running Windows XP right now. An estimated 10% of the U.S. government\u27s computers run Windows XP, including thousands of computers on classified military and diplomatic networks. And the costs of staying put? They will be enormous, too. It turns out that 95% of the world\u27s ATMs are powered by Windows XP, and there is no readily available substitute in the offing. In one example of these exorbitant costs, the price of either extending support or upgrading to another version of Windows for each of Britain\u27s major banks will be in the hundreds of millions. Costs will be similar, or perhaps even higher, worldwide. The failure to continue to patch unpatched computers will also have serious ramifications for society. Hackers will keep scouring Windows XP for flaws, holes, and vulnerabilities for many years after Microsoft ends support for its vaunted operating system. Zero-day vulnerabilities-flaws hackers have long known about but have waited to exploit for fear that the vulnerability will be immediately patched (and could therefore only be used once or for a short time)-will now be exploitable in perpetuity. Experts have repeatedly warned that April 8 could spark a hacker feeding frenzy. Yet, ironically, those who run Windows XP pose a greater threat to others than to themselves. In the special case of governments, enterprise businesses, and financial institutions, failure to patch or migrate will expose the personal data of millions of individuals to theft, fraud, and abuse. When the wave of security breaches from unpatched machines arrives, the lawsuits against companies for failure to secure user data properly will become even more costly. There may also be immense blows to consumer confidence, leading to welfare-reducing market-wide substitutions away from e-commerce

Effect of intracanal time of triple antibiotic paste on its removal from simulated immature roots using passive ultrasonic irrigation and XP-endo Finisher

Background. The aim of this study was to evaluate the effectiveness of different irrigation techniques in the removal of triple antibiotic paste (TAP), which was applied for 7, 21 or 90 days, from simulated root canals of immature teeth. Methods. The root canal spaces of 190 maxillary canine teeth were filled with TAP and randomly divided into a control and 3 experimental groups according to the intracanal medicament period (7, 21 or 90 days). Syringe irrigation (SI), passive ultrasonic irrigation (PUI) and XP-endo Finisher (XP) were used for the removal of TAP (n=20). The amount of remaining medicament was calculated under a stereomicroscope using a 4-grade scoring system. Kruskal-Wallis H and Wilcoxon signedrank tests were used for statistical analyses (P0.05), whereas the efficacy of SI and XP was significantly affected (P0.05); however, at 90-day interval, PUI removed significantly greater amount of TAP than XP did (P<0.05). Conclusion. The time of the TAP in the root canal negatively affected the removal efficacy of the SI and XP-Endo Finisher; however, it did not affect the efficacy of the PUI

DANSSino: a pilot version of the DANSS neutrino detector

DANSSino is a reduced pilot version of a solid-state detector of reactor antineutrinos (to be created within the DANSS project and installed under the industrial 3 GW(th) reactor of the Kalinin Nuclear Power Plant -- KNPP). Numerous tests performed at a distance of 11 m from the reactor core demonstrate operability of the chosen design and reveal the main sources of the background. In spite of its small size (20x20x100 ccm), the pilot detector turned out to be quite sensitive to reactor antineutrinos, detecting about 70 IBD events per day with the signal-to-background ratio about unity.Comment: 16 pages, 11 figures, 3 tables. arXiv admin note: substantial text overlap with arXiv:1304.369

The IBIS soft gamma-ray sky after 1000 INTEGRAL orbits

We report here an all-sky soft gamma-ray source catalog based on IBIS observations performed during the first 1000 orbits of INTEGRAL. The database for the construction of the source list consists of all good quality data available from launch in 2002 up to the end of 2010. This corresponds to $\sim$110 Ms of scientific public observations with a concentrated coverage on the Galactic Plane and extragalactic deep exposures. This new catalog includes 939 sources above a 4.5 sigma significance threshold detected in the 17-100 keV energy band, of which 120 represent previously undiscovered soft gamma-ray emitters. The source positions are determined, mean fluxes are provided in two main energy bands, and are reported together with the overall source exposure. Indicative levels of variability are provided, and outburst times and durations are given for transient sources. Comparison is made with previous IBIS catalogs, and those from other similar missions.Comment: 65 pages, 9 figures, 2 tables, accepted for publication in ApJ Supplement

Inadequate food intake at high temperatures is related to depressed mitochondrial respiratory capacity

Animals, especially ectotherms, are highly sensitive to the temperature of their surrounding environment. Extremely high temperature, for example, induces a decline of average performance of conspecifics within a population, but individual heterogeneity in the ability to cope with elevating temperatures has rarely been studied. In this study, we examined inter-individual variation in feeding ability and consequent growth rate of juvenile brown trout Salmo trutta acclimated to a high temperature (19°C), and investigated the relationship between these metrics of whole-animal performances and among-individual variation in mitochondrial respiration capacity. Food was provided ad libitum yet intake varied ten-fold amongst individuals, resulting in some fish losing weight whilst others continued to grow. Almost half of the variation in food intake was related to variability in mitochondrial capacity: low intake (and hence growth failure) was associated with high leak respiration rates within liver and muscle mitochondria, and a lower coupling of muscle mitochondria. These observations, combined with the inability of fish with low food consumption to increase their intake despite ad libitum food levels, suggest a possible insufficient capacity of the mitochondria for maintaining ATP homeostasis. Individual variation in thermal performance is likely to confer variation in the upper limit of an organism's thermal niche and in turn affect the structure of wild populations in warming environments