Exploring the Experiences and Adaptation Strategies of People Who Develop Work-Related Musculoskeletal Symptoms After Working from Home During the Coronavirus Pandemic

At the beginning of 2020, the COVID-19 pandemic brought many “new ways of life” to people everywhere. The transition from working in an office to working from home forced many individuals to adapt to a new normal. This capstone research study used qualitative case study methodology to provide a detailed look into the challenges and adaptive process of three employees working from home during the COVID-19 pandemic. This capstone study describes the lived experiences of individuals working from home and how they adapted in response to the onset of upper extremity musculoskeletal symptoms. Using the Theory of Occupational Adaptation, the researcher provides insight into the participants’ experiences, occupational challenges, level of dysfunction, and how they adapted, including physical, ergonomic, and psychosocial factors.https://soar.usa.edu/otdcapstones-spring2022/1051/thumbnail.jp

Experimental validation of growth rate normalization relation.

<p><i>E. coli</i> harboring 11 different plasmids driving differential eGFP levels were grown in a single population or in two separate populations. The fitness from the separate populations (represented by blue and red open circles) and combined populations were evaluated and compared. Error bars represent one standard deviation from two independent experiments. The solid black line represents the theoretically ideal relationship between individual and combined fitness.</p

Enrichment ratio of a clone as a function of its abundance in the selected population.

<p>The dynamic range of the method lies between enrichment ratios of-4 to 4 (indicated by horizontal dashed lines) such that (i.) single clones do not dominate the selected population; and (ii.) loss-of-function clones are not completely removed from the population.</p

Errors introduced by different PCR methods.

<p>Identical mutant libraries were prepared for sequencing using three different PCR methods. The number of counts for each library member was compared across the different methods. Each point represents a specific mutant sequence. For each panel, dashed black lines represent the 95% confidence interval for the Poisson noise between different methods. <i>A</i>. Method B v. Method A. Above 10 counts, the data fall almost completely within minimal error predicted by Poisson noise. <i>B</i>. By contrast, Method C shows significant variance in counts relative to Method A.</p

Growth Selection Parameters.

<p>The parameters of growth-based selections should be chosen such that the range of enrichment ratios for the population lies between-4 and 4. <i>A</i>. Enrichment ratios as a function of the individual growth rate compared to the population growth rate. Following one generation of population growth (blue) the enrichment ratios remain around zero. Increasing the number of population generations the experiment is allowed to grow (1 generation, blue, 5 generations, red and 10 generations, green) increases the experimental resolution in discriminating mutant growth phenotypes. <i>B</i>. Enrichment ratios as a function of average population generation (g_p) for various . For values less than one (0.1, red; 0.5, blue; and 0.667, green) the enrichment ratios decrease with increasing population generations. Variants with values values above one (1.5, black) show enhanced enrichment with increasing population generations.</p

Overview of high-resolution sequence-function mapping process.

<p><i>Target Selection</i>. Proteins of interest are selected for interrogation of sequence function relationships. A plasmid containing the gene-encoding sequence is generated. <i>Gene tiling</i>. Starting from this gene sequence, semi-overlapping tiles are generated to cover the entire gene. These tiles are either 150, 250 or 300 bp in length in order to be sequenced in paired-end mode on Illumina deep sequencing platforms. <i>Library Preparation</i>. The single-pot PFunkel method is used to generate a comprehensive single-site saturation mutagenesis library. <i>Selections</i>. Growth-based selections and FACS screens are used to resolve library populations; these selections should not completely converge on a few members of the population. It is important that the initial protein shows activity toward the selection method. <i>Deep Sequencing and Library Preparation</i>. After selection, cells are lysed and plasmid DNA is purified. The specific mutated tile region of the gene of interest is then amplified using overhang PCR, at which time Illumina sequencing primers and adaptors with selection-specific indexes are attached. <i>Data analysis and normalization</i>. Barcoded DNA is sequenced on a standard Illumina platform, analyzed, and normalized using custom scripts. The end result of this analysis is a comprehensive portrait of the effects of sequence on function for thousands of single point mutants in the gene of interest. These portraits can be used for various purposes such as improving protein binding affinity and specificity or improving enzymatic catalytic efficiency.</p

2-step PCR method for deep sequencing preparation of libraries.

<p>PCR reactions are shown for two separate gene tiles containing single mutations (orange and green). Primers are designed to be complementary to flanking regions (grey) of each tile, with encoded single mutations. The first set of primers includes the flanking regions and Illumina sequencing primers (purple). In the next step, outer primers add the Illumina adaptor (pink) and multiplexing index (teal) sequences to the gene. The PCR reaction is performed in a single tube using a 1:2 molar ratio of inner to outer primers and bead purified to remove primer dimer products. The purified library is ready for sequencing without further modifications. While the first set of primers is specific to a single gene tile, the outer primer set is universal.</p