Factors Associated With Consenting to Receive an Automated Email Summary After a Sexual Health Clinic Visit.

BACKGROUND: Electronic health records allow for inexpensive communication with patients. In March 2021, the Melbourne Sexual Health Centre implemented an automated email summary ("Sexual Health Automated Visit Email" [SHAVE]) of a client's visit. This study evaluates the proportion of attendees at a sexual health service who opted in or out of SHAVE. METHODS: This study was conducted at the Melbourne Sexual Health Centre in Australia between March 2021 and June 2022. Univariable and multivariable logistic regression analyses were used to examine the client characteristics associated with consenting to SHAVE. RESULTS: There were 18,528 clients (men, 12,700; women, 5828) included in the final analysis and 55.2% (n = 10,233) consented to receiving SHAVE. Comparing with those who did not have a new sexually transmitted infection diagnosis, clients with a new diagnosis of a sexually transmitted infection, but not HIV, had lower odds of consenting to receiving SHAVE (chlamydia: adjusted odds ratio [aOR], 0.64 [95% confidence interval {CI}, 0.57-0.72]; gonorrhea: aOR, 0.71 [95% CI, 0.62-0.82]; syphilis: aOR, 0.75 [95% CI, 0.59-0.96]). Men had lower odds of consenting when compared with women (men who have sex with women only: aOR, 0.77 [95% CI, 0.71-0.84]; men who have sex with men: aOR, 0.68 [95% CI, 0.62-0.75]). Comparing with those born in Australia or Oceania, clients born in Europe had lower odds of consenting (aOR, 0.81; 95% CI, 0.70-0.94), whereas those born in Latin America or Caribbean had higher odds of consenting (aOR, 1.25; 95% CI, 1.04-1.51). CONCLUSIONS: Email summaries may serve as a valuable strategy to improve health communication and record keeping for clients. Understanding the client characteristics associated with consenting SHAVE will allow for the implementation of strategies to better communicate with clients.

SARS-CoV-2 viral variants can rapidly be identified for clinical decision making and population surveillance using a high-throughput digital droplet PCR assay.

Epidemiologic surveillance of circulating SARS-CoV-2 variants is essential to assess impact on clinical outcomes and vaccine efficacy. Whole genome sequencing (WGS), the gold-standard to identify variants, requires significant infrastructure and expertise. We developed a digital droplet polymerase chain reaction (ddPCR) assay that can rapidly identify circulating variants of concern/interest (VOC/VOI) using variant-specific mutation combinations in the Spike gene. To validate the assay, 800 saliva samples known to be SARS-CoV-2 positive by RT-PCR were used. During the study (July 2020-March 2022) the assay was easily adaptable to identify not only existing circulating VAC/VOI, but all new variants as they evolved. The assay can discriminate nine variants (Alpha, Beta, Gamma, Delta, Eta, Epsilon, Lambda, Mu, and Omicron) and sub-lineages (Delta 417N, Omicron BA.1, BA.2). Sequence analyses confirmed variant type for 124/124 samples tested. This ddPCR assay is an inexpensive, sensitive, high-throughput assay that can easily be adapted as new variants are identified.

Limitations of gene editing assessments in human preimplantation embryos.

Range of DNA repair in response to double-strand breaks induced in human preimplantation embryos remains uncertain due to the complexity of analyzing single- or few-cell samples. Sequencing of such minute DNA input requires a whole genome amplification that can introduce artifacts, including coverage nonuniformity, amplification biases, and allelic dropouts at the target site. We show here that, on average, 26.6% of preexisting heterozygous loci in control single blastomere samples appear as homozygous after whole genome amplification indicative of allelic dropouts. To overcome these limitations, we validate on-target modifications seen in gene edited human embryos in embryonic stem cells. We show that, in addition to frequent indel mutations, biallelic double-strand breaks can also produce large deletions at the target site. Moreover, some embryonic stem cells show copy-neutral loss of heterozygosity at the cleavage site which is likely caused by interallelic gene conversion. However, the frequency of loss of heterozygosity in embryonic stem cells is lower than in blastomeres, suggesting that allelic dropouts is a common whole genome amplification outcome limiting genotyping accuracy in human preimplantation embryos.

The Effect of Resuscitation Residents on the Duration of Pre-induction of Targeted Temperature Management in Out-of-Hospital Cardiac Arrest.

Background The Resuscitation Rotation is a novel second-year emergency medicine rotation focusing on the highest acuity patients, including out-of-hospital cardiac arrest (OHCA). The resuscitation resident (RR) functions as an extra physician during resuscitation and post return of spontaneous circulation (ROSC). The objective of this study is to examine if the presence of a RR decreases the pre-induction interval of targeted temperature management (TTM) for patients following OHCA. Methods A retrospective study was conducted at a tertiary care level 1 trauma center with an annual ED census of 127,323 visits in 2019. We retrospectively reviewed consecutive OHCA patients from September 1, 2014, to July 20, 2020, who underwent TTM. Patients were identified as cases with or without a RR. Clinical characteristics were summarized by the status of RR involvement and compared by using t-test and χ2 test for continuous and categorical variables, respectively. All tests with p < 0.05 were considered to indicate statistical significance. Results Our study population identified 198 adult OHCA patients that underwent TTM from 2014-2020. There were exclusions for missing TTM start time and for missing patient characteristics leaving 176 for final analysis, of which 55 (33.3%) had RR involvement. The mean time (hours) to TTM initiation (ie, the pre-induction phase) for patients involving the RR versus those without was not statistically significant (3.11 vs 3.34, p=0.39). Linear regression analysis indicates that the adjusted effect of RR involvement was not associated with the mean hours of pre-induction (p=0.47). Conclusion There is no statistically significant association of a RR on the duration of the pre-induction phase. Limitations include that both arms had prolonged pre-induction phases. This may represent a non-optimized TTM protocol. Future work will aim to use the RR to improve our pre-induction phase.

Reproductive hormone levels, androgen receptor CAG repeat length and their longitudinal relationships with decline in cognitive subdomains in men: The European Male Ageing Study.

OBJECTIVE: It has been proposed that endogenous sex hormone levels may present a modifiable risk factor for cognitive decline. However, the evidence for effects of sex steroids on cognitive ageing is conflicting. We therefore investigated associations between endogenous hormone levels, androgen receptor CAG repeat length, and cognitive domains including visuoconstructional abilities, visual memory, and processing speed in a large-scale longitudinal study of middle-aged and older men. METHODS: Men aged 40-79 years from the European Male Ageing Study (EMAS) underwent cognitive assessments and measurements of hormone levels at baseline and follow-up (mean = 4.4 years, SD ± 0.3 years). Hormone levels measured included total and calculated free testosterone and estradiol, dihydrotestosterone, luteinizing hormone, follicle-stimulating hormone, dehydroepiandrosterone sulphate and sex hormone-binding globulin. Cognitive function was assessed using the Rey-Osterrieth Complex Figure Copy and Recall, the Camden Topographical Recognition Memory and the Digit Symbol Substitution Test. Multivariate linear regressions were used to examine associations between baseline and change hormone levels, androgen receptor CAG repeat length, and cognitive decline. RESULTS: Statistical analyses included 1,827 and 1,423 participants for models investigating relationships of cognition with hormone levels and CAG repeat length, respectively. In age-adjusted models, we found a significant association of higher baseline free testosterone (ß=-0.001, p=0.005) and dihydrotestosterone levels (ß=-0.065, p=0.003) with greater decline on Rey-Osterrieth Complex Figure Recall over time. However, these effects were no longer significant following adjustment for centre, health, and lifestyle factors. No relationships were observed between any other baseline hormone levels, change in hormone levels, or androgen receptor CAG repeat length with cognitive decline in the measured domains. CONCLUSIONS: In this large-scale prospective study there was no evidence for an association between endogenous sex hormone levels or CAG repeat length and cognitive ageing in men. These data suggest that sex steroid levels do not affect visuospatial function, visual memory, or processing speed in middle-aged and older men.

Frequency-specific Electrocochleography and Traveling Wave Time as a Clinical Test for Menière's Disease.

INTRODUCTION: Menière's disease is a clinical entity with no definitive objective testing. It has been hypothesized that underlying endolymphatic hydrops stiffens the basilar membrane leading to increased speed of the acoustic stimulus, therefore traveling wave velocity has been proposed as an objective test to aid in the diagnosis. The objective of this study is to compare electrocochleography frequency-specific action potential latency, basilar membrane traveling wave time, and summation to action potential (SP/AP) ratio in Menière's and non-Menière's patients. METHODS: Tympanic electrocochleography was performed with frequency-specific action potential latency time and SP/AP ratio recorded. Patient demographics, symptoms, audiogram data, AAO-HNS classification of Menière's disease, management interventions, and follow-up were recorded. Statistical analysis was performed to compare outcome measures across patient groups, demographics, and clinical data. RESULTS: Ninety-one patients (182 ears) were included. There was a significant difference between a "definite" Menière's diagnosis and an "unlikely" or "probable" diagnosis by an average of 13 dB HL for the pure-tone thresholds at 250 Hz on the affected side (p = 0.006). There was no significant difference in pure-tone thresholds at any other frequency, AP latency at any frequency, or AP/SP ratio between the different Menière's classification groups. CONCLUSIONS: Our study fails to show significance of the traveling wave velocity as an objective test for Menière's disease. A significant correlation was found with low-frequency hearing loss between AAO-HNS Menière's classification groups.

Assessing the impact of resuscitation residents on the treatment of cardiopulmonary resuscitation patients.

BACKGROUND: The management of cardiac arrest patients receiving cardiopulmonary resuscitation (CPR) is an essential aspect of emergency medicine (EM) training. At our institution, we have a 1-month Resuscitation Rotation designed to augment resident training in managing critical patients. The objective of this study is to compare 30-day mortality between cardiac arrest patients with resuscitation resident (RR) involvement versus patients without. Our secondary outcome is to determine if RR involvement altered rates of initiating targeted temperature management (TTM). METHODS: This study was conducted at a single site tertiary care Level-1 trauma center with an Emergency Department (ED) census of nearly 130,000 visits per year. Data was collected from 01/01/2015 to 01/01/2018 using electronic medical records via query. Patients admitted with cardiac arrest were separated into two groups, one with RR involvement and one without. Initial rhythm of ventricular fibrillation/tachycardia (VFIB/VTACH), 30-day mortality, history of coronary artery disease (CAD), and initiation of TTM were compared. Statistical analysis was performed. RESULTS: Out of 885 patient encounters, 91 (10.28%) had RR participation. There was no statistical difference in 30-day mortality between patients with RR involvement compared to those without (71.42% vs 66.36%; P = 0.3613). However, TTM was initiated more in the RR group (20.70% vs 8.86%; P = 0.0025). Patients who received TTM also had a lower 30-day mortality compared to those without TTM (52.94% vs 70.87%; P = 0.0020). Patients who were older and had no history of CAD were also noted to have a statistically significant higher 30-day mortality. All other variables were not statistically significant. CONCLUSION: Resuscitation resident involvement with the care of cardiac arrest patients had no impact in 30-day mortality. However, the involvement of RR was associated with a statistically significant increase in the initiation of TTM. One limitation is that RR participated in 10.28% of the cases analyzed herein, thus the two arms are unbalanced in size. Future work may investigate if the increase in TTM in the RR involved cases may portend improved rates of neurologically intact survival or more rapid achievement of goal temperatures.

Elements at the 5' end of Xist harbor SPEN-independent transcriptional antiterminator activity.

The Xist lncRNA requires Repeat A, a conserved RNA element located in its 5' end, to induce gene silencing during X-chromosome inactivation. Intriguingly, Repeat A is also required for production of Xist. While silencing by Repeat A requires the protein SPEN, how Repeat A promotes Xist production remains unclear. We report that in mouse embryonic stem cells, expression of a transgene comprising the first two kilobases of Xist (Xist-2kb) causes transcriptional readthrough of downstream polyadenylation sequences. Readthrough required Repeat A and the ∼750 nucleotides downstream, did not require SPEN, and was attenuated by splicing. Despite associating with SPEN and chromatin, Xist-2kb did not robustly silence transcription, whereas a 5.5-kb Xist transgene robustly silenced transcription and read through its polyadenylation sequence. Longer, spliced Xist transgenes also induced robust silencing yet terminated efficiently. Thus, in contexts examined here, Xist requires sequence elements beyond its first two kilobases to robustly silence transcription, and the 5' end of Xist harbors SPEN-independent transcriptional antiterminator activity that can repress proximal cleavage and polyadenylation. In endogenous contexts, this antiterminator activity may help produce full-length Xist RNA while rendering the Xist locus resistant to silencing by the same repressive complexes that the lncRNA recruits to other genes.

OK Cupid, Stop Bumbling around and Match Me Tinder: Using Dating Apps Across the Life Course.

Social connectedness, sex, and intimacy are all factors associated with positive aging, facing individuals in society across the life course. Phenomenal technological developments in the 21st century have led to the increased use of smartphones, mobile apps, and dating apps for a myriad of services, and engagements. This paper focuses on two specific cohorts' who have the opportunity to engage with dating apps, older adults and young citizens with life-limiting or life-threatening conditions, and highlights issues related to the intersection of technology, societal constructions of age, disability, and online dating.

Circulating TNF-like protein 1A (TL1A) is elevated early in rheumatoid arthritis and depends on TNF.

BACKGROUND: The tumor necrosis factor (TNF) superfamily cytokine TNF-like protein 1A (TL1A) and its receptor DR3 are essential for diverse animal models of autoimmune disease and may be pathogenic in rheumatoid arthritis (RA). However, the relationship of TL1A to disease duration, activity, and response to anti-TNF and other therapies in RA is not clear. METHODS: We measured soluble TL1A in synovial fluid (SF), serum, or plasma from RA first-degree relatives (FDRs) and in early RA and established disease. We measured the effects of anti-TNF and methotrexate (MTX) therapy on circulating TL1A from multiple independent RA treatment trials. We also determined the ability of a blocking anti-TL1A antibody to inhibit clinical disease and articular bone destruction in the murine collagen-induced arthritis (CIA) model of human RA. RESULTS: Soluble TL1A was specifically elevated in the blood and SF of patients with RA compared to patients with other diseases and was elevated early in disease and in at-risk anti-cyclic citrullinated peptide (CCP) (+) first-degree relatives (FDRs). Therapeutic TNF inhibition reduced serum TL1A in both responders and non-responders, whereas TL1A declined following MTX treatment only in responders. In murine CIA, TL1A blockade was clinically efficacious and reduced bone erosions. CONCLUSIONS: TL1A is specifically elevated in RA from early in the disease course and in at-risk FDRs. The decline in TL1A after TNF blockade suggests that TL1A levels may be a useful biomarker for TNF activity in RA. These results support the further investigation of the relationship between TL1A and TNF and TL1A blockade as a potential therapeutic strategy in RA.

Sexual Expression and Cognitive Function: Gender-Divergent Associations in Older Adults.

Prior research demonstrates a positive association between sexual activity and cognitive function in later life. However, the relationship between the type of sexual activity and cognitive function in older adulthood remains unclear. This study explored the associations between the frequency of engaging in different types of sexual activities (intercourse, masturbation, and kissing/petting/fondling) and cognitive function in older women and men. Using data from Wave 6 of the English Longitudinal Study of Aging (ELSA), 1915 women and 2195 men (age range 50-89 years; n = 4110) reporting any type of sexual activity over the past 12 months were included in the study. Multiple regression controlling for age, education, satisfaction with sex life, cohabiting, wealth, general health, physical activity, depression, and loneliness was used to explore the associations between the frequency of engagement in intercourse, masturbation, and kissing/petting/fondling, and two measures of cognitive function: word recall and number sequencing. For women, masturbation was linked to better word recall (p = .008), while for men, kissing/petting/fondling was associated with better number sequencing (p = .035). In women (p = .016) and men (p = .018), dissatisfaction with sex life was associated with better number sequencing. The results point to gendered links between sexual activity and cognitive function. These gender-related divergences may reflect differences in biological/neurological mechanisms, or in cognitive lifestyle factors that could influence cognitive reserve in later life. This novel study underscores the need to delineate the underlying mechanisms of the association between sex and cognition in men and women.

lncRNA-Induced Spread of Polycomb Controlled by Genome Architecture, RNA Abundance, and CpG Island DNA.

Long noncoding RNAs (lncRNAs) cause Polycomb repressive complexes (PRCs) to spread over broad regions of the mammalian genome. We report that in mouse trophoblast stem cells, the Airn and Kcnq1ot1 lncRNAs induce PRC-dependent chromatin modifications over multi-megabase domains. Throughout the Airn-targeted domain, the extent of PRC-dependent modification correlated with intra-nuclear distance to the Airn locus, preexisting genome architecture, and the abundance of Airn itself. Specific CpG islands (CGIs) displayed characteristics indicating that they nucleate the spread of PRCs upon exposure to Airn. Chromatin environments surrounding Xist, Airn, and Kcnq1ot1 suggest common mechanisms of PRC engagement and spreading. Our data indicate that lncRNA potency can be tightly linked to lncRNA abundance and that within lncRNA-targeted domains, PRCs are recruited to CGIs via lncRNA-independent mechanisms. We propose that CGIs that autonomously recruit PRCs interact with lncRNAs and their associated proteins through three-dimensional space to nucleate the spread of PRCs in lncRNA-targeted domains.

A piggyBac-based toolkit for inducible genome editing in mammalian cells.

We describe the development and application of a novel series of vectors that facilitate CRISPR-Cas9-mediated genome editing in mammalian cells, which we call CRISPR-Bac. CRISPR-Bac leverages the piggyBac transposon to randomly insert CRISPR-Cas9 components into mammalian genomes. In CRISPR-Bac, a single piggyBac cargo vector containing a doxycycline-inducible Cas9 or catalytically dead Cas9 (dCas9) variant and a gene conferring resistance to Hygromycin B is cotransfected with a plasmid expressing the piggyBac transposase. A second cargo vector, expressing a single-guide RNA (sgRNA) of interest, the reverse-tetracycline TransActivator (rtTA), and a gene conferring resistance to G418, is also cotransfected. Subsequent selection on Hygromycin B and G418 generates polyclonal cell populations that stably express Cas9, rtTA, and the sgRNA(s) of interest. We show that CRISPR-Bac can be used to knock down proteins of interest, to create targeted genetic deletions with high efficiency, and to activate or repress transcription of protein-coding genes and an imprinted long noncoding RNA. The ratio of sgRNA-to-Cas9-to-transposase can be adjusted in transfections to alter the average number of cargo insertions into the genome. sgRNAs targeting multiple genes can be inserted in a single transfection. CRISPR-Bac is a versatile platform for genome editing that simplifies the generation of mammalian cells that stably express the CRISPR-Cas9 machinery.

RETRACTED: A 5' fragment of Xist can sequester RNA produced from adjacent genes on chromatin.

Xist requires Repeat-A, a protein-binding module in its first two kilobases (2kb), to repress transcription. We report that when expressed as a standalone transcript in mouse embryonic stem cells (ESCs), the first 2kb of Xist (Xist-2kb) does not induce transcriptional silencing. Instead, Xist-2kb sequesters RNA produced from adjacent genes on chromatin. Sequestration does not spread beyond adjacent genes, requires the same sequence elements in Repeat-A that full-length Xist requires to repress transcription and can be induced by lncRNAs with similar sequence composition to Xist-2kb. We did not detect sequestration by full-length Xist, but we did detect it by mutant forms of Xist with attenuated transcriptional silencing capability. Xist-2kb associated with SPEN, a Repeat-A binding protein required for Xist-induced transcriptional silencing, but SPEN was not necessary for sequestration. Thus, when expressed in mouse ESCs, a 5' fragment of Xist that contains Repeat-A sequesters RNA from adjacent genes on chromatin and associates with the silencing factor SPEN, but it does not induce transcriptional silencing. Instead, Xist-induced transcriptional silencing requires synergy between Repeat-A and additional sequence elements in Xist. We propose that sequestration is mechanistically related to the Repeat-A dependent stabilization and tethering of Xist near actively transcribed regions of chromatin.

The dilemma of early preventive oophorectomy in familial small cell carcinoma of the ovary of hypercalcemic type.

•New pathogenic familial SMARCA4 variant, c.3081+1G>T.•Prophylactic surgery in healthy carrier of germline SMARCA4 mutation.•Long term hormone therapy in a 13-year-old girl.

Sleep quality, sleep duration and sexual health among older people: Findings from the English Longitudinal Study of Ageing.

CONTEXT: How sleep quality and sexual health are associated among community-dwelling individuals remains largely unknown. OBJECTIVE: We examined the association of sleep disturbance and sleep duration with a range of measures assessing sexual activities, functioning and concerns in a representative sample of older people. METHODS: Participants were community-dwelling adults aged 50-90+ years from wave 6 (2012/2013) of the English Longitudinal Study of Ageing (ELSA) who reported any sexual activity in the last year. Sleep disturbance, sleep duration and sexual health were measured by self-report at wave 6. Retrospective reports of restless sleep (waves 1-6 [2002-2013]) were also examined. The association between sleep measures and sexual health was assessed using logistic regressions stratified by gender and adjusted for demographic, health and lifestyle factors with results expressed as odds ratios (OR) and 95% confidence intervals (CI). RESULTS: Among both men and women disturbed sleep was associated with reported declines in sexual activity and function over the last year, and increased concern about their sexual desire, frequency of sexual activity and sexual functioning. Robust associations between high sleep disturbance and vaginal pain (OR = 1.67, 95% CI = 1.21, 2.31) and vaginal dryness (OR = 1.69, 95% CI = 1.24, 2.30) were also observed among women. Retrospective reports of restless sleep showed a dose-response relationship with reported declines in sexual health over the last year, and increased concerns about sexual expression and functioning. CONCLUSIONS: Self-reported sleep disturbance and retrospective restless sleep were mainly associated with subjective assessments of recent declines in sexual activity and functioning, and higher levels of sexual concerns.

Functional classification of long non-coding RNAs by k-mer content.

The functions of most long non-coding RNAs (lncRNAs) are unknown. In contrast to proteins, lncRNAs with similar functions often lack linear sequence homology; thus, the identification of function in one lncRNA rarely informs the identification of function in others. We developed a sequence comparison method to deconstruct linear sequence relationships in lncRNAs and evaluate similarity based on the abundance of short motifs called k-mers. We found that lncRNAs of related function often had similar k-mer profiles despite lacking linear homology, and that k-mer profiles correlated with protein binding to lncRNAs and with their subcellular localization. Using a novel assay to quantify Xist-like regulatory potential, we directly demonstrated that evolutionarily unrelated lncRNAs can encode similar function through different spatial arrangements of related sequence motifs. K-mer-based classification is a powerful approach to detect recurrent relationships between sequence and function in lncRNAs.