Role of the Srs2-Rad51 Interaction Domain in Crossover Control in Saccharomyces cerevisiae.

Saccharomyces cerevisiae Srs2, in addition to its well-documented antirecombination activity, has been proposed to play a role in promoting synthesis-dependent strand annealing (SDSA). Here we report the identification and characterization of an SRS2 mutant with a single amino acid substitution (srs2-F891A) that specifically affects the Srs2 pro-SDSA function. This residue is located within the Srs2-Rad51 interaction domain and embedded within a protein sequence resembling a BRC repeat motif. The srs2-F891A mutation leads to a complete loss of interaction with Rad51 as measured through yeast two-hybrid analysis and a partial loss of interaction as determined through protein pull-down assays with purified Srs2, Srs2-F891A, and Rad51 proteins. Even though previous work has shown that internal deletions of the Srs2-Rad51 interaction domain block Srs2 antirecombination activity in vitro, the Srs2-F891A mutant protein, despite its weakened interaction with Rad51, exhibits no measurable defect in antirecombination activity in vitro or in vivo Surprisingly, srs2-F891A shows a robust shift from noncrossover to crossover repair products in a plasmid-based gap repair assay, but not in an ectopic physical recombination assay. Our findings suggest that the Srs2 C-terminal Rad51 interaction domain is more complex than previously thought, containing multiple interaction sites with unique effects on Srs2 activity

Fowler-Nordheim-like local injection of photoelectrons from a silicon tip

Tunneling between a photo-excited p-type silicon tip and a gold surface is studied as a function of tip bias, tip/sample distance and light intensity. In order to extend the range of application of future spin injection experiments, the measurements are carried out under nitrogen gas at room temperature. It is found that while tunneling of valence band electrons is described by a standard process between the semiconductor valence band and the metal, the tunneling of photoelectrons obeys a Fowler-Nordheim-like process directly from the conduction band. In the latter case, the bias dependence of the photocurrent as a function of distance is in agreement with theoretical predictions which include image charge effects. Quantitative analysis of the bias dependence of the dark and photocurrent spectra gives reasonable values for the distance, and for the tip and metal work functions. For small distances image charge effects induce a vanishing of the barrier and the bias dependence of the photocurrent is exponential. In common with many works on field emission, fluctuations in the tunneling currents are observed. These are mainly attributed to changes in the prefactor for the tunneling photocurrent, which we suggest is caused by an electric-field-induced modification of the thickness of the natural oxide layer covering the tip apex.Comment: 12 pages, 11 figures. Submitted to Phys. Rev.

Conclusions and Future Research Directions

In this book, we have discussed channel estimation for various situations in PLNC, including frequency flat fading environment, frequency selective fading environment, and time-selective fading environment. In each environment, we demonstrated how the channel estimation is different from the conventional point-to-point transmission as well as from the uni-directional relay network. The key idea is that the individual channel knowledge should be obtained at three nodes, i.e., the terminals and the relay, within one round of the data exchange. One may, of course, apply more complicated training process, say, training each channel link separately and share the information through feedback channels but such processing is not compatible with the overall structure of the data frame. Moreover, we developed channel estimation algorithms that fit the speciality of different environments, for example in frequency selective fading environment it is possible to remove the redundant estimates so that the overall training length could be reduced, while in time selective fading environment the individual BEM coefficient is estimated instead of the channel parameters

Application of machine learning algorithms to the study of noise artifacts in gravitational-wave data

The sensitivity of searches for astrophysical transients in data from the Laser Interferometer Gravitational-wave Observatory (LIGO) is generally limited by the presence of transient, non-Gaussian noise artifacts, which occur at a high enough rate such that accidental coincidence across multiple detectors is non-negligible. These “glitches” can easily be mistaken for transient gravitational-wave signals, and their robust identification and removal will help any search for astrophysical gravitational waves. We apply machine-learning algorithms (MLAs) to the problem, using data from auxiliary channels within the LIGO detectors that monitor degrees of freedom unaffected by astrophysical signals. Noise sources may produce artifacts in these auxiliary channels as well as the gravitational-wave channel. The number of auxiliary-channel parameters describing these disturbances may also be extremely large; high dimensionality is an area where MLAs are particularly well suited. We demonstrate the feasibility and applicability of three different MLAs: artificial neural networks, support vector machines, and random forests. These classifiers identify and remove a substantial fraction of the glitches present in two different data sets: four weeks of LIGO’s fourth science run and one week of LIGO’s sixth science run. We observe that all three algorithms agree on which events are glitches to within 10% for the sixth-science-run data, and support this by showing that the different optimization criteria used by each classifier generate the same decision surface, based on a likelihood-ratio statistic. Furthermore, we find that all classifiers obtain similar performance to the benchmark algorithm, the ordered veto list, which is optimized to detect pairwise correlations between transients in LIGO auxiliary channels and glitches in the gravitational-wave data. This suggests that most of the useful information currently extracted from the auxiliary channels is already described by this model. Future performance gains are thus likely to involve additional sources of information, rather than improvements in the classification algorithms themselves. We discuss several plausible sources of such new information as well as the ways of propagating it through the classifiers into gravitational-wave searches

Machiavelli Meets Michelangelo

This article examines newspaper coverage of the arts in Singapore to consider the role of the island state’s newspapers in the development and documentation of Singapore’s growing arts scene. Sampling two constructed weeks for each of 10 years, 1999 to 2008, content analysis is used to examine arts coverage in the Straits Times and Lianhe Zaobao. The study benefits from groundwork laid by Janssen’s 1999 study of arts coverage in Dutch newspapers, in which not only quantity of content was reported but also hierarchical attention was paid to art forms over decades. Singapore is of interest as it represents a country where neither the arts nor newspapers are declining, and both enjoy significant overseeing by the government

Genetically modified semisynthetic bioluminescent photoprotein variants: simultaneous dual-analyte assay in a single well employing time resolution of decay kinetics

Progress in the miniaturization and automation of complex analytical processes depends largely on increasing the sensitivity, diversity, and robustness of current labels. Because of their ubiquity and ease of use, fluorescent, enzymatic, and bioluminescent labels are often employed in such miniaturized and multiplexed formats, with each type of label having its own unique advantages and drawbacks. The ultrasensitive detection limits of bioluminescent reporters are especially advantageous when dealing with very small sample volumes and biological fluids. However, bioluminescent reporters currently do not have the multiplexing capability that fluorescent labels do. In an effort to address this limitation, we have developed a method of discriminating two semisynthetic aequorin variants from one another using time resolution. In this work we paired two aequorin conjugates with different coelenterazine analogues and then resolved the two signals from one another using the difference in decay kinetics and half-life times. Utilizing this time-resolution, we then developed a simultaneous, dual-analyte, single well assay for 6-keto-prostaglandin-FI-alpha and angiotensin II, two important cardiovascular molecules

Role of the Srs2-Rad51 Interaction Domain in Crossover Control in Saccharomyces cerevisiae.

Saccharomyces cerevisiae Srs2, in addition to its well-documented antirecombination activity, has been proposed to play a role in promoting synthesis-dependent strand annealing (SDSA). Here we report the identification and characterization of an SRS2 mutant with a single amino acid substitution (srs2-F891A) that specifically affects the Srs2 pro-SDSA function. This residue is located within the Srs2-Rad51 interaction domain and embedded within a protein sequence resembling a BRC repeat motif. The srs2-F891A mutation leads to a complete loss of interaction with Rad51 as measured through yeast two-hybrid analysis and a partial loss of interaction as determined through protein pull-down assays with purified Srs2, Srs2-F891A, and Rad51 proteins. Even though previous work has shown that internal deletions of the Srs2-Rad51 interaction domain block Srs2 antirecombination activity in vitro, the Srs2-F891A mutant protein, despite its weakened interaction with Rad51, exhibits no measurable defect in antirecombination activity in vitro or in vivo Surprisingly, srs2-F891A shows a robust shift from noncrossover to crossover repair products in a plasmid-based gap repair assay, but not in an ectopic physical recombination assay. Our findings suggest that the Srs2 C-terminal Rad51 interaction domain is more complex than previously thought, containing multiple interaction sites with unique effects on Srs2 activity

Trabecular bone score is associated with volumetric bone density and microarchitecture as assessed by central QCT and HRpQCT in Chinese American and white women.

Although high-resolution peripheral quantitative computed tomography (HRpQCT) and central quantitative computed tomography (QCT) studies have shown bone structural differences between Chinese American (CH) and white (WH) women, these techniques are not readily available in the clinical setting. The trabecular bone score (TBS) estimates trabecular microarchitecture from dual-energy X-ray absorptiometry spine images. We assessed TBS in CH and WH women and investigated whether TBS is associated with QCT and HRpQCT indices. Areal bone mineral density (aBMD) by dual-energy X-ray absorptiometry, lumbar spine (LS) TBS, QCT of the LS and hip, and HRpQCT of the radius and tibia were performed in 71 pre- (37 WH and 34 CH) and 44 postmenopausal (21 WH and 23 CH) women. TBS did not differ by race in either pre- or postmenopausal women. In the entire cohort, TBS positively correlated with LS trabecular volumetric bone mineral density (vBMD) (r = 0.664), femoral neck integral (r = 0.651), trabecular (r = 0.641) and cortical vBMD (r = 0.346), and cortical thickness (C/I; r = 0.540) by QCT (p < 0.001 for all). TBS also correlated with integral (r = 0.643), trabecular (r = 0.574) and cortical vBMD (r = 0.491), and C/I (r = 0.541) at the total hip (p < 0.001 for all). The combination of TBS and LS aBMD predicted more of the variance in QCT measures than aBMD alone. TBS was associated with all HRpQCT indices (r = 0.20-0.52) except radial cortical thickness and tibial trabecular thickness. Significant associations between TBS and measures of HRpQCT and QCT in WH and CH pre- and postmenopausal women demonstrated here suggest that TBS may be a useful adjunct to aBMD for assessing bone quality