Attitudes toward COVID-19 vaccination among urogynecology patients.

Objective: Urogynecology patients skew older and often require intimate exams to treat non-life-threatening conditions, thus making care particularly susceptible to the effects of COVID-19. We aim to understand COVID-19 vaccination attitudes amongst urogynecology patients during the pandemic, which has adversely affected healthcare delivery, to identify measures that can be undertaken to improve care going forward. Study design: Cross-sectional surveys of urogynecology patients were conducted in three different states (Delaware, New Jersey, and Pennsylvania) between March and August 2021. Demographics, vaccination status, and COVID-19 attitudes were analyzed. Chi-square tests investigated group differences between vaccine-hesitant and unhesitant subjects. Results: Out of 158 surveys, eighty-three percent of respondents were vaccinated or planning to get vaccinated. Older age and regular flu vaccination predicted COVID-19 vaccination. Ninety-three percent of subjects were comfortable attending in-person visits. However, vaccine-hesitant individuals were less comfortable attending in-person and were significantly less likely to be comforted by clinic interventions, such as masking. Hesitant and unhesitant groups trusted doctors or medical professionals most for their source of COVID-19 information. Conclusions: Urogynecology patients mirror the general older population's COVID-19 vaccination attitudes, with distinct differences between hesitant and unhesitant groups. Vaccine-hesitant subjects appear less comfortable coming to in-person visits and less comforted by possible interventions, posing a difficult obstacle for clinics to overcome. High levels of patient trust suggest healthcare providers can play a crucial role in encouraging COVID-19 vaccination and combating misinformation.

Quality of Pelvic Floor Therapy Videos on YouTube: Does Popularity Predict Reliability?

ABSTRACT: This study reviewed instructional videos on YouTube regarding pelvic floor physical therapy and assessed the association between the videos' popularity and the reliability of the videos' content. YouTube was searched using the terms relevant to pelvic floor muscle training. The first 100 videos for each search term were screened, and relevant metrics were collected for those meeting the inclusion criteria. Videos were assessed by 2 independent, trained health care professionals for reliability using the Medical Quality Video Evaluation (MQ-VET) tool, the modified DISCERN tool, the Journal of American Medical Association benchmark criteria, and the Global Quality Score. Popularity was assessed using the Video Power Index (VPI). Higher values for all scoring systems correlated with greater reliability and greater popularity, respectively. Five hundred videos were screened. Two hundred thirty-four videos were duplicates, and 99 did not meet the inclusion criteria. A total of 167 videos were reviewed. The median VPI and MQ-VET score was 201,114.76 (interquartile range, 7,194,020.29) and 48.00 (interquartile range, 12.75), respectively. Spearman's R value was 0.292 (P < 0.001), demonstrating a weak positive correlation between MQ-VET scores and VPI. The interrater reliability of the MQ-VET was good, with an intraclass correlation coefficient of 0.86 (95% confidence interval, 0.71-0.92). In summary, we identified a statistically significant but weak positive correlation between the reliability and popularity of YouTube videos about pelvic floor physical therapy.

Bisulfite treatment and single-molecule real-time sequencing reveal D-loop length, position, and distribution.

Displacement loops (D-loops) are signature intermediates formed during homologous recombination. Numerous factors regulate D-loop formation and disruption, thereby influencing crucial aspects of DNA repair, including donor choice and the possibility of crossover outcome. While D-loop detection methods exist, it is currently unfeasible to assess the relationship between D-loop editors and D-loop characteristics such as length and position. Here, we developed a novel in vitro assay to characterize the length and position of individual D-loops with near base-pair resolution and deep coverage, while also revealing their distribution in a population. Non-denaturing bisulfite treatment modifies the cytosines on the displaced strand of the D-loop to uracil, leaving a permanent signature for the displaced strand. Subsequent single-molecule real-time sequencing uncovers the cytosine conversion patch as a D-loop footprint. The D-loop Mapping Assay is widely applicable with different substrates and donor types and can be used to study factors that influence D-loop properties.

Rdh54/Tid1 inhibits Rad51-Rad54-mediated D-loop formation and limits D-loop length.

Displacement loops (D-loops) are critical intermediates formed during homologous recombination. Rdh54 (a.k.a. Tid1), a Rad54 paralog in Saccharomyces cerevisiae, is well-known for its role with Dmc1 recombinase during meiotic recombination. Yet contrary to Dmc1, Rdh54/Tid1 is also present in somatic cells where its function is less understood. While Rdh54/Tid1 enhances the Rad51 DNA strand invasion activity in vitro, it is unclear how it interplays with Rad54. Here, we show that Rdh54/Tid1 inhibits D-loop formation by Rad51 and Rad54 in an ATPase-independent manner. Using a novel D-loop Mapping Assay, we further demonstrate that Rdh54/Tid1 uniquely restricts the length of Rad51-Rad54-mediated D-loops. The alterations in D-loop properties appear to be important for cell survival and mating-type switch in haploid yeast. We propose that Rdh54/Tid1 and Rad54 compete for potential binding sites within the Rad51 filament, where Rdh54/Tid1 acts as a physical roadblock to Rad54 translocation, limiting D-loop formation and D-loop length.

Arginine methylation augments Sbp1 function in translation repression and decapping.

The fate of messenger RNA in cytoplasm plays a crucial role in various cellular processes. However, the mechanisms that decide whether mRNA will be translated, degraded or stored remain unclear. Single stranded nucleic acid binding protein (Sbp1), an Arginine-Glycine-Glycine (RGG-motif) protein, is known to promote transition of mRNA into a repressed state by binding eukaryotic translation initiation factor 4G1 (eIF4G1) and to promote mRNA decapping, perhaps by modulation of Dcp1/2 activity. Sbp1 is known to be methylated on arginine residues in RGG-motif; however, the functional relevance of this modification in vivo remains unknown. Here, we report that Sbp1 is arginine-methylated in an hnRNP methyl transferase (Hmt1)-dependent manner and that methylation is enhanced upon glucose deprivation. Characterization of an arginine-methylation-defective (AMD) mutant provided evidence that methylation affects Sbp1 function in vivo. The AMD mutant is compromised in causing growth defect upon overexpression, and the mutant is defective in both localizing to and inducing granule formation. Importantly, the Sbp1-eIF4G1 interaction is compromised both for the AMD mutant and in the absence of Hmt1. Upon overexpression, wild-type Sbp1 increases localization of another RGG motif containing protein, Scd6 (suppressor of clathrin deficiency) to granules; however, this property of Sbp1 is compromised in the AMD mutant and in the absence of Hmt1, indicating that Sbp1 repression activity could involve other RGG-motif translation repressors. Additionally, the AMD mutant fails to increase localization of the decapping activator DEAD box helicase homolog to foci and fails to rescue the decapping defect of a dcp1-2Δski8 strain, highlighting the role of Sbp1 methylation in decapping. Taken together, these results suggest that arginine methylation modulates Sbp1 role in mRNA fate determination.

Dynamic Processing of Displacement Loops during Recombinational DNA Repair.

Displacement loops (D-loops) are pivotal intermediates of homologous recombination (HR), a universal DNA double strand break (DSB) repair pathway. We developed a versatile assay for the physical detection of D-loops in vivo, which enabled studying the kinetics of their formation and defining the activities controlling their metabolism. Nascent D-loops are detected within 2 h of DSB formation and extended in a delayed fashion in a genetic system designed to preclude downstream repair steps. The majority of nascent D-loops are disrupted by two pathways: one supported by the Srs2 helicase and the other by the Mph1 helicase and the Sgs1-Top3-Rmi1 helicase-topoisomerase complex. Both pathways operate without significant overlap and are delineated by the Rad54 paralog Rdh54 in an ATPase-independent fashion. This study uncovers a layer of quality control of HR relying on nascent D-loop dynamics.

Homologous recombination and the repair of DNA double-strand breaks.

Homologous recombination enables the cell to access and copy intact DNA sequence information in trans, particularly to repair DNA damage affecting both strands of the double helix. Here, we discuss the DNA transactions and enzymatic activities required for this elegantly orchestrated process in the context of the repair of DNA double-strand breaks in somatic cells. This includes homology search, DNA strand invasion, repair DNA synthesis, and restoration of intact chromosomes. Aspects of DNA topology affecting individual steps are highlighted. Overall, recombination is a dynamic pathway with multiple metastable and reversible intermediates designed to achieve DNA repair with high fidelity.

Arginine methylation promotes translation repression activity of eIF4G-binding protein, Scd6.

Regulation of translation plays a critical role in determining mRNA fate. A new role was recently reported for a subset of RGG-motif proteins in repressing translation initiation by binding eIF4G1. However the signaling mechanism(s) that leads to spatial and temporal regulation of repression activity of RGG-motif proteins remains unknown. Here we report the role of arginine methylation in regulation of repression activity of Scd6, a conserved RGG-motif protein. We demonstrate that Scd6 gets arginine methylated at its RGG-motif and Hmt1 plays an important role in its methylation. We identify specific methylated arginine residues in the Scd6 RGG-motif in vivo We provide evidence that methylation augments Scd6 repression activity. Arginine methylation defective (AMD) mutant of Scd6 rescues the growth defect caused by overexpression of Scd6, a feature of translation repressors in general. Live-cell imaging of the AMD mutant revealed that it is defective in inducing formation of stress granules. Live-cell imaging and pull-down results indicate that it fails to bind eIF4G1 efficiently. Consistent with these results, a strain lacking Hmt1 is also defective in Scd6-eIF4G1 interaction. Our results establish that arginine methylation augments Scd6 repression activity by promoting eIF4G1-binding. We propose that arginine methylation of translation repressors with RGG-motif could be a general modulator of their repression activity.