Recombinant viruses described in this study were generated using transformation-associated recombination in yeast as we described previously77. SARS-CoV-2 recombinant viruses carrying the Delta or Omicron variant spike within the ancestral WT lineage B backbone were assembled by transformation-associated recombination in yeast using a set of relevant overlapping complementary DNA fragments to assemble the modified genomes. RNA transcribed in vitro from the recombinant genomes was used to rescue the viruses following transfection into BHK cells stably expressing ACE2 and SARS-CoV-2 N protein. Two clones of each rescued virus were passaged (P1) into VERO E6 cells and their genomes verified by sequencing using Oxford Nanopore as described above73.
the passage justin cronin pdf 82
For any analysis, estimates of causal effects would be strengthened with data showing how CAP laws actually affected gun storage behaviors, but national longitudinal data on firearm storage patterns are limited. The researchers who have been able to leverage national data on firearm storage practices and child gun access have found that CAP laws are associated with higher rates of safe storage (Prickett, Martin-Storey, and Crosnoe, 2014) and lower rates of gun carrying among high school students (Anderson and Sabia, 2018). However, others have found that significant estimates of higher safe storage rates among firearm owners in CAP states are rendered small and imprecise in models that adjust for individual-level and state-level differences (Miller et al., 2022); this possibly indicates that associations found between CAP laws and safe storage could be attributable to other correlated factors, such as individual-level characteristics (e.g., the number and types of firearms owned by survey respondents) or state-level characteristics (e.g., overall prevalence of household gun ownership). Cross-sectional analyses like this one cannot establish, however, whether storage practices improved after passage of a CAP law. Ideally, matching of gun owners in states with and without the law would occur prior to the implementation of CAP laws to observe how they affected storage behavior.
Some have further questioned whether knowledge of CAP laws is sufficiently well understood by gun owners to account for the apparent effects of these laws. If a person is unaware of a law, it is difficult to associate her or his behavior with that law. It may be, however, that knowledge of the law is not the key driver of improvements in gun storage after a CAP law is passed. If, for instance, passage of the law changes discourse around safe storage among gun owners and their influencers, the law could affect more gun owners than just those who know about the law. Too little work has been done on the mechanisms by which CAP laws or other laws influence their measured outcomes to conclude that low public awareness of the laws proves that they cannot be effective.
As with any lexicon-based approach, however, it is possible that our dictionary might be capturing something more or less than nostalgia (so-called false positives). Is the more frequent appearance of these words truly indicative of "nostalgia," or perhaps something else entirely, or maybe just semantic noise? As a second step, we used the process of machine learning to try to predict the number of nostalgic passages per novel in our collection. Here, we identified 300 nostalgic passages by hand, where "passage" was defined as a unit of 1,000 words of text and "nostalgic" was defined as "any scene that includes a retrospective looking back at one's own life, ideally in a longing kind of way."20
2ff7e9595c