Targeted Mutagenesis of the Oligopeptide Repeat Domain of the Yeast Prion Sup35

The formation of prions in the baker’s yeast Saccharomyces cerevisiae is determined by amino acid composition rather than the primary sequence of amino acids. The infectious amyloid proteins known as prions undergo nucleation and propagation, two distinct activities critical for prion formation. The ability for prions to be transferred from cell to cell, or propagate, is of interest not only in yeast prions but also in prion diseases such as the mammalian spongiform encephalopathies. Prion formation has been widely studied in yeast prions, however, the fundamental mechanisms behind the specific process of propagation of prions from cell to cell are not yet understood. In the most well-studied yeast prion, the prion form [PSI+] of Sup35, a domain of 5 ½ degenerate oligopeptide repeats called the oligopeptide repeat domain (ORD) has been shown to be important for prion propagation and to have a distinct amino acid composition as compared to the nucleation domain region. A library mutagenesis experiment has identified amino acids that favor or disfavor prion propagation in yeast cells. To confirm the results of the random library mutagenesis experiment, we generated several clones in which a portion of the ORD (the fourth oligopeptide repeat) was replaced with defined sequences expected to propagate or fail to propagate

Legal origin and the evolution of environmental quality

We extend the empirical literature on the environmental Kuznets curve (EKC) by showing the legal origin matters for the evolution of environmental quality. Using observations of ambient sulfur dioxide levels, we find that the EKC for French and British legal origin countries diverge as incomes rise, with the EKC for French legal origin countries lying significantly below that for countries of British legal origin. This finding is robust to the inclusion of proxies for democracy and corruption, the institutional variables emphasized in the current EKC literature. Our results are consistent with the idea that the British common law tradition places a greater emphasis on private relative to collective property rights.Environmental quality, institutions, legal origin. economic development, evironmental Kuznets curve

Composing For A Singer: The Vocal Qualities of Adriana Ferrarese del Bene Exemplified Through Four Mozart Arias

Adriana Ferrarese del Bene played Susanna in the 1789 revival of Le Nozze di Figaro in Vienna, for which Mozart rewrote both arias sung by the character. She later became the soprano who premiered the role of Fiordiligi in Così fan tutte. In both instances, Mozart carefully considered the vocal abilities of Ferrarese when composing the arias for Susanna and Fiordiligi, including Ferrarese\u27s powerful chest voice, ability to negotiate large leaps, and agility. Through melodic analysis of the four arias Mozart wrote specifically for her voice, an understanding of Mozart\u27s ideal sound is apparent. Musical examples from each of the four arias are included to demonstrate specific moments in which Mozart highlight\u27s Ferrarese\u27s strengths

Mapping the Beta-Sheet Structure of the Yeast Prion Sup35 through Creation of Targeted Mutant Forms

Proteins with an aggregated form rich in beta-sheet structure are known as amyloids, of which a subset are infectious. These infectious proteins are known as prions and cause diseases including bovine spongiform encephalopathy (“Mad Cow” disease). Several prions have been identified in the baker’s yeast, Saccharomyces cerevisiae. One of the most well-studied yeast prions is the protein Sup35. To understand the fine protein structure of Sup35 better, we used PCR-based mutagenesis to introduce a lysine residue (a charged amino acid) at five defined places in the protein sequence of Sup35. We describe our process for creating these mutant versions and the results of DNA sequencing of each mutant version. The next step will be to assess prion formation and stability of clones with the correct sequences. Understanding the behavior of yeast prions has proven helpful in understanding human amyloid diseases and further studies on these yeast prions, including Sup35, will expand our knowledge further

Social Media: A Good Alternative, for Alternative Service of Process

A cost efficient and effective means of alternative service of process in civil and commercial cases upon both domestic and international defendants lies in social media. Traditional methods of service of process are preferred, but are not always practicable. When a defendant cannot be reached through traditional methods, judges should use their discretion and allow alternative service by social media in appropriate cases. Current methods of alternative service, such as publication, are not efficient or effective. By allowing alternative service of process via social media in certain cases, the defendant is much more likely to receive actual notice in a cost-effective manner

Evaluating the Recovery of DNA from Adhesive Tape after Exposure to Heat and Humidity: Assessing the Degradation Index and STR Profile

It is not uncommon for forensic DNA samples to be degraded or of low quantity; these types of samples can pose challenges throughout analysis and an informative STR profile is not guaranteed. DNA analysis is also very expensive and timely; therefore, it is necessary to understand where to sample a piece of evidence to obtain the best quality and quantity of DNA from the substrate, especially in the case of trace DNA samples which may be difficult to detect. It is also necessary to know when to proceed with analysis given the quality of the sample. The aim of this research was to evaluate the effect of adhesive tape on the degradation of DNA when exposed to heat and humidity and to assess the usefulness of the QuantifilerTM Trio degradation index to estimate the level of degradation in an STR profile. DNA was deposited on or between layers of duct tape or electrical tape and incubated in a humidity chamber for up to two weeks. The degradation index and profile slope were determined for each sample to assess whether the type of tape or number of layers had a significant effect on the degradation of the DNA sample. Multiple linear regression was also used to assess the relationship between the degradation index, amplification input, and the profile slope. The type of tape, number of layers of tape, and treatment length were found to have a significant effect on the degradation index of DNA samples, however, the treatment length was the only factor that had a significant effect on the slope of the DNA profile. These results indicate that the type of tape and number of layers may significantly affect the degradation of a DNA sample given a long enough treatment period, however the degradation index alone cannot be used to evaluate the level of degradation in a DNA sample or estimate the slope of a DNA profile; factors such as the quantity of DNA amplified and method of degradation affect the profile slope as well

Molecular characterization of ALDRXv4, an aldose reductase orthologue isolated from xerophyta viscosa, in response to abiotic stress

Includes bibliographical references (leaves 61-70)