Reduced mutation rate and increased transformability of transposon-free Acinetobacter baylyi ADP1-ISx

ABSTRACT The genomes of most bacteria contain mobile DNA elements that can contribute to undesirable genetic instability in engineered cells. In particular, transposable insertion sequence (IS) elements can rapidly inactivate genes that are important for a designed function. We deleted all six copies of IS 1236 from the genome of the naturally transformable bacterium Acinetobacter baylyi ADP1. The natural competence of ADP1 made it possible to rapidly repair deleterious point mutations that arose during strain construction. In the resulting ADP1-ISx strain, the rates of mutations inactivating a reporter gene were reduced by 7- to 21-fold. This reduction was higher than expected from the incidence of new IS 1236 insertions found during a 300-day mutation accumulation experiment with wild-type ADP1 that was used to estimate spontaneous mutation rates in the strain. The extra improvement appears to be due in part to eliminating large deletions caused by IS 1236 activity, as the point mutation rate was unchanged in ADP1-ISx. Deletion of an error-prone polymerase ( dinP ) and a DNA damage response regulator ( umuD Ab [the umuD gene of A. baylyi ]) from the ADP1-ISx genome did not further reduce mutation rates. Surprisingly, ADP1-ISx exhibited increased transformability. This improvement may be due to less autolysis and aggregation of the engineered cells than of the wild type. Thus, deleting IS elements from the ADP1 genome led to a greater than expected increase in evolutionary reliability and unexpectedly enhanced other key strain properties, as has been observed for other clean-genome bacterial strains. ADP1-ISx is an improved chassis for metabolic engineering and other applications. IMPORTANCE Acinetobacter baylyi ADP1 has been proposed as a next-generation bacterial host for synthetic biology and genome engineering due to its ability to efficiently take up DNA from its environment during normal growth. We deleted transposable elements that are capable of copying themselves, inserting into other genes, and thereby inactivating them from the ADP1 genome. The resulting “clean-genome” ADP1-ISx strain exhibited larger reductions in the rates of inactivating mutations than expected from spontaneous mutation rates measured via whole-genome sequencing of lineages evolved under relaxed selection. Surprisingly, we also found that IS element activity reduces transformability and is a major cause of cell aggregation and death in wild-type ADP1 grown under normal laboratory conditions. More generally, our results demonstrate that domesticating a bacterial genome by removing mobile DNA elements that have accumulated during evolution in the wild can have unanticipated benefits. </jats:p

Crosslinking of aromatic polyamides via pendant propargyl groups

Methods for crosslinking N-methyl substituted aromatic polyamides were investigated in an effort to improve the applicability of these polymers as matrix resins for Kavlar trademark fiber composites. High molecular weight polymers were prepared from isophthaloyl dichloride and 4,4'- bis(methylamino)diphenylmethane with varying proportions of the N,N'bispropargyl diamine incorporated as a crosslinking agent. The propargylcontaining diamines were crosslinked thermally and characterized by infrared spectroscopy, differential scanning calorimetry, and thermogravimetric analysis. Attempts were also made to crosslink polyamide films by exposure to ultraviolet light, electron beam, and gamma radiation

Mask exposure during COVID-19 changes emotional face processing

Faces are one of the key ways that we obtain social information about others. They allow people to identify individuals, understand conversational cues, and make judgements about others’ mental states. When the COVID-19 pandemic hit the United States, widespread mask-wearing practices were implemented, causing a shift in the way Americans typically interact. This introduction of masks into social exchanges posed a potential challenge—how would people make these important inferences about others when a large source of information was no longer available? We conducted two studies that investigated the impact of mask exposure on emotion perception. In particular, we measured how participants used facial landmarks (visual cues) and the expressed valence and arousal (affective cues), to make similarity judgements about pairs of emotion faces. Study 1 found that in August 2020, participants with higher levels of mask exposure used cues from the eyes to a greater extent when judging emotion similarity than participants with less mask exposure. Study 2 measured participants’ emotion perception in both April and September 2020 –before and after widespread mask adoption—in the same group of participants to examine changes in the use of facial cues over time. Results revealed an overall increase in the use of visual cues from April to September. Further, as mask exposure increased, people with the most social interaction showed the largest increase in the use of visual facial cues. These results provide evidence that a shift has occurred in how people process faces such that the more people are interacting with others that are wearing masks, the more they have learned to focus on visual cues from the eye area of the face

Plant Community Assessment and Management Recommendations for Minneapolis Park Natural Areas

In 2017 the Minneapolis Park and Recreation Board (MPRB) began a two-phase study to collect quantitative and qualitative data for urban park natural areas in Minneapolis, MN parks to inform management activities. The first phase took existing GIS data and quality ranking systems and tailored them to the Minneapolis park system. The second phase, which is still in process, involves field checking the data, applying the quality ranking system, and writing a management plan

Definitive observation of the dark triplet ground state of charged excitons in high magnetic fields

The ground state of negatively charged excitons (trions) in high magnetic fields is shown to be a dark triplet state, confirming long-standing theoretical predictions. Photoluminescence (PL), reflection, and PL excitation spectroscopy of CdTe quantum wells reveal that the dark triplet trion has lower energy than the singlet trion above 24 Tesla. The singlet-triplet crossover is "hidden" (i.e., the spectral lines themselves do not cross due to different Zeeman energies), but is confirmed by temperature-dependent PL above and below 24 T. The data also show two bright triplet states.Comment: 4 figure

Managing a Multi-Forage Rotational Stocking Unit without Farming Equipment and with the Addition of Economical Fringe Benefits

A specialized introduced forage rotational stocking grazing unit was managed without usual grassland farming equipment and with various alternative management and production techniques as fringe benefits. The forage production techniques have been successful for many years. Livestock production and economic perimeters have been average to above average. The greatest sold beef yield has been 713 pounds of beef per acre. That is excellent considering the low-capital overhead management style of the unit

The effect of pregabalin or duloxetine on arthritis pain: a clinical and mechanistic study in people with hand osteoarthritis (vol 10, pg 2437, 2017)

Sofat N, Harrison A, Russell MD, et al. J Pain Res. 2017;10:2437&ndash;2449.On page 2443, Table 3, Placebo column, NRS section, the difference was reported as: &ndash;0.9 (&ndash;0.2 to 0.2). This is incorrect, and it should read as follows: &ndash;0.9 (&ndash;2.0 to 0.2).Read the original articl