A scalable and cost-efficient rRNA depletion approach to enrich RNAs for molecular biology investigations.

Transcriptomics analyses play pivotal roles in understanding the complex regulatory networks that govern cellular processes. The abundance of rRNAs, which account for 80%-90% of total RNA in eukaryotes, limits the detection and investigation of other transcripts. While mRNAs and long noncoding RNAs have poly(A) tails that are often used for positive selection, investigations of poly(A)- RNAs, such as circular RNAs, histone mRNAs, and small RNAs, typically require the removal of the abundant rRNAs for enrichment. Current approaches to deplete rRNAs for downstream molecular biology investigations are hampered by restrictive RNA input masses and high costs. To address these challenges, we developed rRNA Removal by RNaseH (rRRR), a method to efficiently deplete rRNAs from a wide range of human, mouse, and rat RNA inputs and of varying qualities at a cost 10- to 20-fold cheaper than other approaches. We used probe-based hybridization and enzymatic digestion to selectively target and remove rRNA molecules while preserving the integrity of non-rRNA transcripts. Comparison of rRRR to two commercially available approaches showed similar rRNA depletion efficiencies and comparable off-target effects. Our developed method provides researchers with a valuable tool for investigating gene expression and regulatory mechanisms across a wide range of biological systems at an affordable price that increases the accessibility for researchers to enter the field, ultimately advancing our understanding of cellular processes.

Comparing the characteristics of people living with and without HIV in long-term care and home care in Ontario, Canada.

Population aging and successful drug therapy in human immunodeficiency virus (HIV) management mean that more people are living longer with HIV. As these individuals age, they become more at risk of developing other chronic health conditions which will have many implications for disease management and choice of care setting. As people living with HIV turn to home care and long-term care (LTC) settings for care, understanding the particular needs of this population is becoming increasingly important. This study sought to describe the sociodemographic, clinical, and social attributes of people living with HIV in the home care and institutional environments. This work involved secondary analysis of data collected from both the international Resident Assessment Instruments (interRAI) home care and minimum data set instruments in the Canadian province of Ontario. Descriptive analysis was used to describe key attributes of people living with and without HIV in LTC, complex continuing care, and home care settings. A comparison of differences between people living with HIV across the three environments was also done using Chi-square analysis. People living with HIV were often younger, male and unmarried than other populations in the care settings studied. Together with specific health needs associated with issues like mental health and social isolation, people living with HIV represent a population with complex and distinctive health needs. Finding ways to better understand the needs of this vulnerable population will help to develop strategies to provide better formal and informal care and improve the quality of life of this group. interRAI standardized assessment instruments may be important tools for meeting this challenge.

The Family Education Diabetes Series: improving health in an urban-dwelling American Indian community.

Community-based participatory research has shown great promise as a mutually engaging and respectful way to partner contemporary biomedical knowledge with the lived experience, wisdom, and customs of American Indian people. Designed and implemented through this approach, our Family Education Diabetes Series (FEDS) has evidenced pilot and longitudinal physiological data supporting its effectiveness. However, the multifaceted nature of the program makes it difficult to know which factors are responsible for its success. This difficulty hinders efforts to improve the FEDS and/or inform others' work to advance similar projects. In this study, we conducted a qualitative investigation using talking circles to explore participants' views about what elements of the FEDS are most salient. Our findings suggest that social support and group-oriented sequences hold the most value. We conclude that an emphasis on these processes (instead of program content per se) is most indicated in effecting behavior change and facilitating ongoing disease management.

Impact of chronic obstructive pulmonary disease on survival and neurologic outcomes in adults with in-hospital cardiac arrest.

BACKGROUND: Little data exists regarding the association of chronic obstructive pulmonary disease (COPD) on outcomes in the setting of in-hospital cardiac arrest (IHCA). We sought to assess the impact of COPD on mortality and neurologic outcomes in adults with IHCA. METHODS: The study population included 593 consecutive hospitalized patients with IHCA undergoing ACLS-guided resuscitation at an academic tertiary medical center from 2012-2018. The primary and secondary outcomes of interest were survival to discharge and favorable neurological outcome (defined as a Glasgow Outcome Score of 4-5) respectively. RESULTS: Of the 593 patients studied, 162 (27.3%) had COPD while 431 (72.7%) did not. Patients with COPD were older, more often female, and had higher Charlson Comorbidity score. Location of cardiac arrest, initial rhythm, duration of cardiopulmonary resuscitation, and rates of defibrillation and return of spontaneous circulation were similar in both groups. Patients with COPD had significantly lower rates of survival to discharge (10.5% vs 21.6%, p = 0.002) and favorable neurologic outcomes (7.4% vs 15.9%, p = 0.007). In multivariable analyses, COPD was independently associated with lower rates of survival to discharge [odds ratio (OR) 0.54, 95% confidence interval (CI) 0.30-0.98, p = 0.041]. CONCLUSIONS: In this contemporary prospective registry of adults with IHCA, COPD was independently associated with significantly lower rates of survival to discharge.

Proteasome ß5 subunit overexpression improves proteostasis during aging and extends lifespan in Drosophila melanogaster.

The ß5 subunit of the proteasome has been shown in worms and in human cell lines to be regulatory. In these models, ß5 overexpression results in upregulation of the entire proteasome complex which is sufficient to increase proteotoxic stress resistance, improve metabolic parameters, and increase longevity. However, fundamental questions remain unanswered, including the temporal requirements for ß5 overexpression and whether ß5 overexpression can extend lifespan in other species. To determine if adult-only overexpression of the ß5 subunit can increase proteasome activity in a different model, we characterized phenotypes associated with ß5 overexpression in Drosophila melanogaster adults. We find that adult-only overexpression of the ß5 subunit does not result in transcriptional upregulation of the other subunits of the proteasome as they do in nematodes and human cell culture. Despite this lack of a regulatory role, boosting ß5 expression increases the chymotrypsin-like activity associated with the proteasome, reduces both the size and number of ubiquitinated protein aggregates in aged flies, and increases longevity. Surprisingly, these phenotypes were not associated with increased resistance to acute proteotoxic insults or improved metabolic parameters.