Integrated multi-omics analyses identify anti-viral host factors and pathways controlling SARS-CoV-2 infection.

Host anti-viral factors are essential for controlling SARS-CoV-2 infection but remain largely unknown due to the biases of previous large-scale studies toward pro-viral host factors. To fill in this knowledge gap, we perform a genome-wide CRISPR dropout screen and integrate analyses of the multi-omics data of the CRISPR screen, genome-wide association studies, single-cell RNA-Seq, and host-virus proteins or protein/RNA interactome. This study uncovers many host factors that are currently underappreciated, including the components of V-ATPases, ESCRT, and N-glycosylation pathways that modulate viral entry and/or replication. The cohesin complex is also identified as an anti-viral pathway, suggesting an important role of three-dimensional chromatin organization in mediating host-viral interaction. Furthermore, we discover another anti-viral regulator KLF5, a transcriptional factor involved in sphingolipid metabolism, which is up-regulated, and harbors genetic variations linked to COVID-19 patients with severe symptoms. Anti-viral effects of three identified candidates (DAZAP2/VTA1/KLF5) are confirmed individually. Molecular characterization of DAZAP2/VTA1/KLF5-knockout cells highlights the involvement of genes related to the coagulation system in determining the severity of COVID-19. Together, our results provide further resources for understanding the host anti-viral network during SARS-CoV-2 infection and may help develop new countermeasure strategies.

Faslpr gene dosage tunes the extent of lymphoproliferation and T cell differentiation in lupus.

Sle1 and Faslpr are two lupus susceptibility loci that lead to manifestations of systemic lupus erythematosus. To evaluate the dosage effects of Faslpr in determining cellular and serological phenotypes associated with lupus, we developed a new C57BL/6 (B6) congenic lupus strain, B6.Sle1/Sle1.Faslpr/+ (Sle1homo.lprhet) and compared it with B6.Faslpr/lpr (lprhomo), B6.Sle1/Sle1 (Sle1homo), and B6.Sle1/Sle1.Faslpr/lpr (Sle1homo.lprhomo) strains. Whereas Sle1homo.lprhomo mice exhibited profound lymphoproliferation and early mortality, Sle1homo.lprhet mice had a lifespan comparable to B6 mice, with no evidence of splenomegaly or lymphadenopathy. Compared to B6 monogenic lupus strains, Sle1homo.lprhet mice exhibited significantly elevated serum ANA antibodies and increased proteinuria. Additionally, Sle1homo.lprhet T cells had an increased propensity to differentiate into Th1 cells. Gene dose effects of Faslpr were noted in upregulating serum IL-1⍺, IL-2, and IL-27. Taken together, Sle1homo.lprhet strain is a new C57BL/6-based model of lupus, ideal for genetic studies, autoantibody repertoire investigation, and for exploring Th1 effector cell skewing without early-age lymphoproliferative autoimmunity.

Patterns of contraceptive use through later reproductive years: A cohort study of Australian women with chronic disease.

BACKGROUND: Pregnancies among women with chronic disease are associated with poor maternal and fetal outcomes. There is a need to understand how women use or don't use contraception across their reproductive years to better inform the development of preconception care strategies to reduce high risk unintended pregnancies, including among women of older reproductive age. However, there is a lack of high-quality longitudinal evidence to inform such strategies. We examined patterns of contraceptive use among a population-based cohort of reproductive aged women and investigated how chronic disease influenced contraceptive use over time. METHODS AND FINDINGS: Contraceptive patterns from 8,030 women of reproductive age from the Australian Longitudinal Study on Women's Health (1973-78 cohort), who were at potential risk of an unintended pregnancy were identified using latent transition analysis. Multinomial mixed-effect logistic regression models were used to evaluate the relationship between contraceptive combinations and chronic disease. Contraception non-use increased between 2006 and 2018 but was similar between women with and without chronic disease (13.6% vs. 12.7% among women aged 40-45 years in 2018). When specific contraceptive use patterns were examined over time, differences were found for women with autoinflammatory diseases only. These women had increased odds of using condom and natural methods (OR = 1.20, 95% CI = 1.00, 1.44), and sterilisation and other methods (OR = 1.61, 95% CI = 1.08, 2.39) or no contraception (OR = 1.32, 95% CI = 1.04, 1.66), compared to women without chronic disease using short-acting methods and condoms. CONCLUSION: Potential gaps in the provision of appropriate contraceptive access and care exist for women with chronic disease, particularly for women diagnosed with autoinflammatory conditions. Development of national guidelines as well as a clear coordinated contraceptive strategy that begins in adolescence and is regularly reviewed during care management through their main reproductive years and into perimenopause is required to increase support for, and agency among, women with chronic disease.

Impact of medication reviews on potentially inappropriate medications and associated costs among older women in aged care.

BACKGROUND: The Residential Medication Management Review (RMMR) service is a large investment by the Australian government and involves considerable time commitment. OBJECTIVES: This study determined the impact of RMMRs on the use of potentially inappropriate medications (PIMs), benzodiazepines and antidepressants, described patterns of PIM use, and examined costs. METHODS: The study included 5576 participants from the Australian Longitudinal Study on Women's Health from 2005 to 2017. Three generalised estimating equation (GEE) models were specified to separately analyse the impact of RMMRs on the use of PIMs, benzodiazepines and antidepressants. Descriptive statistics were used to present, at each year, the proportions of participants with PIMs, patterns of PIMs and costs of PIMs. RESULTS: There was no evidence for an association between the use of RMMRs and the use of PIMs (OR = 0.99; 95% CI = 0.88, 1.11), benzodiazepines (OR = 1.02; 95% CI = 0.95, 1.08) or antidepressants (OR = 0.99; 95% CI = 0.90, 1.10) in the following year. There were few differences in the use of particular classes of PIMs, nor any differences in the median benefits paid by government or out-of-pocket costs, between participants who did and did not receive RMMRs. There was a slight increase in median OOP costs and a decrease in government benefits over time. CONCLUSIONS: There was a lack of long-term changes on use of PIMs, however, its appropriate use must be considered during RMMRs. Healthcare professionals have an obligation to optimise the service to reduce medication costs whilst improving health outcomes among individuals residing in RACF.

Patterns of contraceptive use among young Australian women with chronic disease: findings from a prospective cohort study.

BACKGROUND: Given chronic disease is increasing among young women and unintended pregnancies among these women are associated with poor maternal and fetal outcomes, these women would benefit from effective preconception care. However, there is a lack of understanding of how these women use or don't use contraception to inform such interventions. This study examined patterns of contraceptive use among an Australian cohort of young women and investigated the influence of chronic disease on contraceptive use over time. METHODS: Using data from 15,244 young women from the Australian Longitudinal Study on Women's Health (born 1989-1995), latent transition analysis was performed to identify distinct contraceptive patterns among women who were at risk of an unintended pregnancy. Multinomial mixed-effect models were used to evaluate the relationship between contraceptive combinations and chronic disease. RESULTS: Contraceptive use for women with cardiac and autoinflammatory diseases differed to women without chronic disease over the observation period. Compared to women without chronic disease using the pill, women with cardiac disease had double the odds of using 'other' contraception and condoms (OR = 2.20, 95% CI 1.34, 3.59) and a modest increase in the odds of using the combined oral contraceptive pill and condoms (OR = 1.39, 95% CI 1.03, 1.89). Compared to women without chronic disease who used the pill, women with autoinflammatory disease had increased odds of using LARC and condoms (OR = 1.58, 95% CI 1.04, 2.41), using 'other' contraception and condoms (OR = 1.69, 95% CI 1.11, 2.57), and using the combined oral contraceptive pill and condoms (OR = 1.38, 95% CI 1.09, 1.75). No differences in contraceptive patterns over the observation period were found for women with asthma or diabetes when compared to women without chronic disease. CONCLUSION: The findings identified a need for effective contraceptive counselling as part of routine chronic disease care and improved communication between health care providers and women with chronic disease to improve young women's contraceptive knowledge and agency in contraceptive choice, particularly for those with cardiac or autoinflammatory conditions. This may be the key to reducing high-risk unintended pregnancies among this vulnerable population.

Chronic disease is increasing among young women and unintended pregnancies among these women are associated with poor outcomes for both the mother and baby. To optimise outcomes, it is important for these women to plan pregnancies and use effective contraception until such time. However, there is a lack of understanding of how these women use or don't use contraception, particularly with respect to highly effective contraception. This study examined patterns of contraceptive use among an Australian cohort of young women (born 1989­1995) and investigated the influence of chronic disease on contraceptive use over time. We found differences in contraceptive use over time for women with cardiac disease and those with autoinflammatory diseases. Importantly, compared to women without chronic disease using the pill alone, women with cardiac disease had double the odds of using low efficacy contraception. While women with autoinflammatory disease were 69% more likely to use long-acting methods combined with condoms, these women were also 70% more likely to use low efficacy contraception, compared to women without chronic disease who used the pill only. Contraceptive patterns did not differ for women with asthma or diabetes from women without chronic disease. The findings identified a need for effective contraceptive counselling as part of routine chronic disease care and improved communication between health care providers and women with chronic disease to improve young women's contraceptive knowledge and contraceptive decision-making, particularly for those with cardiac or autoinflammatory conditions. This may be the key to reducing high-risk unintended pregnancies among this vulnerable population.

Decreased in-hospital mortality rate following implementation of a comprehensive electronic medical record system.

AIM: To evaluate changes in in-hospital mortality rate following implementation of a comprehensive electronic medical record (EMR) system. METHODS: Before and after study of 355,709 hospital discharges, over an 8-year period, at a paediatric teaching hospital. The major outcome measures were crude number of in-hospital deaths, deaths per 1000 discharges, and standardised mortality ratio. RESULTS: Primary analysis of data from 2 years before and 2 years after EMR go-live showed a reduction in absolute mortality of 33 deaths, a reduction in the mortality rate of 0.48 per 1000 discharges (95% CI 0.09, 0.88 per 1000): and a relative 22% decrease (95% CI: 4%, 36%, P = 0.02) in deaths per 1000 discharges from 2.20 to 1.72. There was also a reduction in standardised mortality ratio of 47% (95% CI: 18%, 66%, P = 0.004). Post-hoc analysis of mortality rates for an additional 2-year pre-intervention period indicated that these changes in the mortality rate were not part of a pre-existing downward trend. Further analysis of an additional 20-month post-intervention period suggests that the reduced mortality rate has been sustained. CONCLUSION: We documented evidence of a clinically important decrease in in-hospital mortality rate following the implementation of a modern comprehensive EMR system in an Australian paediatric teaching hospital. The study does not prove a causal relationship, and it is possible that other factors explain some, or all, of this difference, but no changes in the hospital population or other major interventions were identified as alternative explanations for this observed change.

Increases in use of Medicare Benefits Schedule mental health items among women who gave birth in New South Wales, 2009-2015.

OBJECTIVE: To report rates of Medicare Benefits Schedule (MBS) mental health item use among a sample of women who gave birth in NSW (2009-2015) and examine if the SAFE START policy increased use of these items among perinatal women. METHODS: Data was drawn from women participating in the Australian Longitudinal Study on Women's Health 1973-1978 cohort, linked to data from the NSW Perinatal Data Collection and MBS. RESULTS: Use of Medicare-subsidised mental health items increased 2.7-fold among perinatal women (n=1,453) between 2009 and 2015 (4.1% versus 11.0% respectively), compared to a 1.3-fold increase among non-perinatal women (n=1,800, 6.3% versus 8.4% respectively). However, the increased use of MBS mental health items among perinatal women was not observed to be impacted by the SAFE START policy, after accounting for time trends. CONCLUSION: There was a substantial increase in the use of MBS mental health items among women in NSW between 2009 and 2015, with a more pronounced increase among women who had given birth compared to those who had not. Implications for public health: This study provides important information about changes in mental health service use during a time of significant investment in perinatal mental health, and demonstrates the value of longitudinal survey data linked with administrative health data to evaluate the impact of health policy.

The association between birth weight and proxy-reported health-related quality of life among children aged 5 - 10 years old: A linked data analysis.

BACKGROUND: Birth weight has a substantial effect on children's cognitive development, physical capability, and emotional development, which in turn impact on Health-Related Quality of Life (HRQoL). Generally, evidence indicates that children born with low birth weight tend to have poorer proxy-reported HRQoL, particularly at school age. However, there is limited evidence on whether variation in HRQoL exists across the entire range of possible birth weights. This study aimed to examine the association between birth weight and proxy-reported HRQoL among children aged 5-10 years old. METHODS: Data from the 1973-78 cohort of the Australian Longitudinal Study on Women's Health were linked with state-based Perinatal Data Collections and the Mothers and their Children's Health study for 1,589 mothers and 2,092 children aged 5 - 10 years old. Generalized estimating equations were used to model the association between birth weight and proxy-reported HRQoL measured by the Pediatric Quality of Life Inventory 4.0. Results are presented as odds ratios with 95 % confidence intervals. RESULTS: In this study, 15.61 % of children were at risk of impaired proxy-reported HRQoL. Each 100-gram increase in birth weight was associated with a 3 % reduction in the odds of impaired HRQoL (AOR = 0.97; 95 % CI: 0.94, 0.99). However, there was only limited evidence of an effect within the normal birth weight range (AOR = 0.97; 95 % CI: 0.94, 1.01). CONCLUSIONS: The findings indicate that increased birth weight was protective against impaired HRQoL, although there was limited evidence of variability within the normal birth weight range. This study contributes to the existing literature by not only emphasizing the impact of low birth weight on children's health and health-related outcomes but also by focusing on the variability within the normal birth weight range, particularly in a setting where low birth weight is less prevalent.

Increased chronic disease prevalence among the younger generation: Findings from a population-based data linkage study to inform chronic disease ascertainment among reproductive-aged Australian women.

BACKGROUND: Chronic disease represents an ongoing public health challenge in Australia with women disproportionately affected and at younger ages compared to men. Accurate prevalence and ascertainment of chronic disease among women of reproductive age at the population level is essential for meeting the family planning and reproductive health challenges that chronic diseases pose. This study estimated the prevalence of chronic disease among younger Australian women of reproductive age, in order to ascertain key conditions that would benefit from targeted family planning support strategies. METHODS AND FINDINGS: Population-level survey data from the 1973-78 and 1989-95 cohorts of the Australian Longitudinal Study on Women's Health were linked to health service use, pharmaceutical, cancer and cause of death data to ascertain the prevalence and chronic disease trends for ten chronic health conditions associated with poor maternal and foetal outcomes. Individual chronic disease algorithms were developed for each chronic disease of interest using the available linked datasets. Lifetime prevalence of chronic disease varied substantially based on each individual data source for each of the conditions of interest. When all data sources were considered, all conditions with the exception of mental health conditions were higher among women in the 1973-78 cohort. However, when focused on point prevalence at similar ages (approximately 25-30 years), the chronic disease trend for women in the 1989-95 cohort was substantially higher, particularly for mental health conditions (70.4% vs 23.6%), diabetes (4.5% vs 1.3%) and multimorbidity (17.9% vs 9.1%). CONCLUSIONS: Given the low concordance between individual data sources, the use of multiple data sources are recommended for chronic disease research focused on women of reproductive age. In order to reduce the increasing chronic disease and multimorbidity trend among women, strategic chronic disease interventions are required to be implemented in childhood and adolescence to ensure the long-term health of not only current but also future generations.

Contraceptive use among women through their later reproductive years: Findings from an Australian prospective cohort study.

OBJECTIVE: Examine patterns of contraceptive use and contraceptive transitions over time among an Australian cohort of women through their later reproductive years. STUDY DESIGN: Latent Transition Analysis was performed using data on 8,197 women from the Australian Longitudinal Study on Women's Health's 1973-78 cohort to identify distinct patterns of contraceptive use across 2006, 2012 and 2018. Women were excluded from the analysis at time points where they were not at risk of an unintended pregnancy. Latent status membership probabilities, item-response probabilities, transitions probabilities and the effect of predictors on latent status membership were estimated and reported. RESULTS: Patterns of contraceptive use were relatively consistent over time, particularly for high efficacy contraceptive methods with 71% of women using long-acting reversible contraceptives in 2012 also using long-acting reversible contraceptives in 2018. Multiple contraceptive use was highest in 2006 when women were aged 28-33 years (19.3%) but declined over time to 14.3% in 2018 when women were aged 40-45 years. Overall, contraceptive patterns stabilised as the women moved into their late 30s and early 40s. CONCLUSIONS: Although fertility declines with age, the stability of contraceptive choice and continued use of short-acting contraception among some women suggests that a contraceptive review may be helpful for women during perimenopause so that they are provided with contraceptive options most appropriate to their specific circumstances.

Home Medicines Review and frailty among community-dwelling older women.

OBJECTIVES: Home Medicines Reviews (HMRs) can optimize medications for frail older adults. This study aimed to determine the use of HMRs according to frailty status and the association between frailty and use of HMRs. METHODS: The study included 9139 female participants enrolled in the Australian Longitudinal Study on Women's Health from 2003 (aged 77-82 years) to 2017 (aged 91-96 years). Generalized estimating equations (GEEs) using log-binomial regressions were used to determine associations using repeated measures on individuals over time. KEY FINDINGS: The majority of participants in the study remained non-frail and did not receive HMRs from 2003 [7116 (77.86%)] to 2017 [1240 (71.31%)]. The use of HMRs was low in both groups with 33 (1.68%; 95% CI, 1.16 to 2.36) frail and 64 (0.89%; 95% CI, 0.69 to 1.14) non-frail participants receiving HMRs in 2003; by 2017, 19 (4.19%; 95% CI, 2.54 to 6.46) frail and 45 (3.50%; 95% CI, 2.57 to 4.66) non-frail participants received HMRs. Frailty was not associated with receiving a HMR (RR 1.06; 95% CI, 0.95 to 1.20), although for every 1-year increase, participants were 10% more likely to receive a HMR (RR 1.10; 95% CI, 1.09 to 1.11). Participants with continuous polypharmacy, ≥4 chronic diseases, >4 general practitioner visits and Department of Veterans Affairs coverage were more likely to receive a HMR. CONCLUSIONS: Despite the proven value of HMRs for frail older people, HMRs were not used for most frail and non-frail community-dwelling women in this study. Reasons for low use of the service should be explored, with interventions to raise awareness of the benefits of the service.

The Association between the Five-Minute Apgar Score and Neurodevelopmental Outcomes among Children Aged 8-66 Months in Australia.

This study aimed to evaluate the association of the five-minute Apgar score and neurodevelopmental outcomes in children by taking the entire range of Apgar scores into account. Data from the Australian Longitudinal Study of Women's Health (ALSWH) and Mothers and their Children's Health (MatCH) study were linked with Australian state-based Perinatal Data Collections (PDCs) for 809 children aged 8-66 months old. Generalized estimating equations were used to model the association between the five-minute Apgar scores and neurodevelopmental outcomes, using STATA software V.15. Of the 809 children, 614 (75.3%) had a five-minute Apgar score of 9, and 130 (16.1%) had an Apgar score of 10. Approximately 1.9% and 6.2% had Apgar scores of 0-6 and 7-8, respectively. Sixty-nine (8.5%) of children had a neurodevelopmental delay. Children with an Apgar score of 0-6 (AOR = 5.7; 95% CI: 1.2, 27.8) and 7-8 (AOR = 4.1; 95% CI: 1.2, 14.1) had greater odds of gross-motor neurodevelopment delay compared to children with an Apgar score of 10. Further, when continuously modelled, the five-minute Apgar score was inversely associated with neurodevelopmental delay (AOR = 0.75; 95% CI: 0.60, 0.93). Five-minute Apgar score was independently and inversely associated with a neurodevelopmental delay, and the risks were higher even within an Apgar score of 7-8. Hence, the Apgar score may need to be taken into account when evaluating neurodevelopmental outcomes in children.

Residential Medication Management Reviews and continuous polypharmacy among older Australian women.

Background Polypharmacy is an important consideration for the provision of Residential Medication Management Reviews (RMMRs) among older women given their enhanced risk of medication-related problems and admission to residential aged care (RAC). Objectives To determine the prevalence of the use of RMMRs among older women in RAC, and the association between RMMRs and polypharmacy, medications, and costs. Setting Older Australian women aged 79-84 years in 2005 who had at least one Medicare Benefits Schedule and Pharmaceutical Benefits Scheme record, received a service in aged care, and consented to data linkage. Methods Generalised estimating equations were used to determine the association between polypharmacy and RMMRs, while adjusting for confounding variables. Main outcome measures Prevalence of the use of RMMRs among older women in RAC, association between RMMRs and polypharmacy, medications, and costs. Results Most participants did not have continuous polypharmacy and did not receive RMMRs from 2005 [451 (67.4%)] until 2017 [666 (66.6%)]. Participants with continuous polypharmacy were 17% more likely to receive a RMMR (risk ratio 1.17; 95% confidence interval 1.11, 1.25). Participants in their final year of life and residing in outer regional/remote/very remote Australia were less likely to receive RMMRs. Out-of-pocket medication costs increased over time, and alendronate and aspirin were common contributors to polypharmacy among participants who received RMMRs. Conclusion Polypharmacy was associated with receiving RMMRs and around two-thirds of women who are entitled to a RMMR never received one. There is potential to improve the use of medicines by increasing awareness of the service among eligible individuals, their carers and health care professionals.

Determinants of neonatal near miss in Australia: A multilevel analysis.

BACKGROUND: Neonatal Near Miss (NNM) is a situation where a newborn narrowly survived the neonatal period. It has been hypothesized that identifying factors that contribute to the occurrence of NNM and taking timely interventions could enhance the quality of newborn care. However, there is limited evidence in Australia. This study aimed to identify the determinants of NNM in Australia. METHODS: Data from the 1973-78 cohort of the Australian Longitudinal Study on Women's Health (ALSWH) were linked with state-based Perinatal Data Collections (PDC) for 3655 mothers and 5526 newborns who were born between 01 January 2007 and 31 December 2015. A newborn was considered as a near miss case if presented with any of the pragmatic criteria (gestational age <32 weeks, birth weight <1500 g, five-minute Apgar score <7) and survived the neonatal period. A multilevel multivariable logistic regression model was used to identify the determinants of NNM. RESULTS: Of the total 5526 live births included in this study, 95 live births met the criteria for NNM, corresponding to an incidence of 17.2 per 1000 live births. After controlling for potential confounders, maternal age 31-34 years (AOR = 2.57; 95% CI: 1.05, 6.30) and 35 years and above (AOR = 4.03; 95% CI: 1.58, 10.31), caesarean section (AOR = 2.24; 95% CI: 1.09, 4.57), and gestational hypertension (AOR = 2.63; 95% CI: 1.21, 5.71) increased the odds of NNM. CONCLUSION: Inclusion of NNM evaluations into newborn care and early screening and interventions for women who become pregnant at older age and those with pregnancy complications could improve the quality of newborn care and reduce neonatal morbidity.

Prevalence and association of continuous polypharmacy and frailty among older women: A longitudinal analysis over 15 years.

OBJECTIVES: This study aimed to determine the prevalence of continuous polypharmacy and hyperpolypharmacy, determine medications that contribute to continuous polypharmacy, and examine the association between frailty and continuous polypharmacy. STUDY DESIGN: A prospective study using data from the Australian Longitudinal Study on Women's Health. Women aged 77-82 years in 2003, and 91-96 years in 2017 were analysed, linking the Pharmaceutical Benefits Scheme data to participants' survey data. MAIN OUTCOME MEASURES: The association between frailty and continuous polypharmacy was determined using generalised estimating equations for log binomial regressions, controlling for confounding variables. Descriptive statistics were used to determine the proportion of women with polypharmacy, and medications that contributed to polypharmacy. RESULTS: The proportion of women with continuous polypharmacy increased over time as they aged. Among participants who were frail (n = 833) in 2017, 35.9 % had continuous polypharmacy and 1.32 % had hyperpolypharmacy. Among those who were non-frail (n = 1966), 28.2 % had continuous polypharmacy, and 1.42 % had hyperpolypharmacy. Analgesics (e.g. paracetamol) and cardiovascular medications (e.g. furosemide and statins) commonly contributed to continuous polypharmacy among frail and non-frail women. Accounting for time and other characteristics, frail women had an 8% increased risk of continuous polypharmacy (RR 1.08; 95 % CI 1.05, 1.11) compared to non-frail women. CONCLUSIONS: Combined, polypharmacy and frailty are key clinical and public health challenges. Given that one-third of women had continuous polypharmacy, monitoring and review of medication use among older women are important, and particularly among women who are frail.

Integrating genome-wide CRISPR immune screen with multi-omic clinical data reveals distinct classes of tumor intrinsic immune regulators.

BACKGROUND: Despite approval of immunotherapy for a wide range of cancers, the majority of patients fail to respond to immunotherapy or relapse following initial response. These failures may be attributed to immunosuppressive mechanisms co-opted by tumor cells. However, it is challenging to use conventional methods to systematically evaluate the potential of tumor intrinsic factors to act as immune regulators in patients with cancer. METHODS: To identify immunosuppressive mechanisms in non-responders to cancer immunotherapy in an unbiased manner, we performed genome-wide CRISPR immune screens and integrated our results with multi-omics clinical data to evaluate the role of tumor intrinsic factors in regulating two rate-limiting steps of cancer immunotherapy, namely, T cell tumor infiltration and T cell-mediated tumor killing. RESULTS: Our studies revealed two distinct types of immune resistance regulators and demonstrated their potential as therapeutic targets to improve the efficacy of immunotherapy. Among them, PRMT1 and RIPK1 were identified as a dual immune resistance regulator and a cytotoxicity resistance regulator, respectively. Although the magnitude varied between different types of immunotherapy, genetically targeting PRMT1 and RIPK1 sensitized tumors to T-cell killing and anti-PD-1/OX40 treatment. Interestingly, a RIPK1-specific inhibitor enhanced the antitumor activity of T cell-based and anti-OX40 therapy, despite limited impact on T cell tumor infiltration. CONCLUSIONS: Collectively, the data provide a rich resource of novel targets for rational immuno-oncology combinations.

Frailty and potentially inappropriate medications using the 2019 Beers Criteria: findings from the Australian Longitudinal Study on Women's Health (ALSWH).

BACKGROUND: Frailty is an essential consideration with potentially inappropriate medications (PIMs), especially among older women. AIMS: This study determined the use of potentially inappropriate medications according to frailty status using the Beers Criteria 2019, identified medications that should be flagged as potentially inappropriate and harmful depending on individual health factors, and determined the association between frailty and PIMs, adjusted for characteristics associated with PIMs. METHODS: This prospective longitudinal study included 9355 participants aged 77-82 years at baseline (2003). Frailty was measured using the FRAIL (fatigue, resistance, ambulation, illness and loss of weight) scale. Generalised estimating equations using log-binomial regressions determined the association between frailty and risk of using PIMs. RESULTS: Among participants who were frail and non-frail at baseline, the majority used ≥ 3 PIMs (74.2% and 58.5%, respectively). At 2017, the proportion using ≥ 3 PIMs remained constant in the frail group (72.0%) but increased in the non-frail group (66.0%). Commonly prescribed medications that may be potentially inappropriate in both groups included benzodiazepines, proton-pump inhibitors and non-steroidal anti-inflammatory drugs, and risperidone was an additional contributor in the non-frail group. When adjusted for other characteristics, frail women had a 2% higher risk of using PIMs (RR 1.02; 95% CI 1.01, 1.03). CONCLUSION: Given that the majority of frail women were using medications that may have been potentially inappropriate, it is important to consider both frailty and PIMs as indicators of health outcomes, and to review the need for PIMs for women aged 77-96 years who are frail.

Common combinations of medications used among oldest-old women: a population-based study over 15 years.

BACKGROUND: Older people use many medications, but combinations of medications used among the oldest old (≥ 80 years) are not commonly reported. AIMS: This study aimed to determine common combinations of medications used among women aged 77-96 years and to describe characteristics associated with these combinations. METHODS: A cohort study of older women enroled in the Australian Longitudinal Study on Women's Health over a 15-year period was used to determine combinations of medications using latent class analysis. Multinomial logistic regression was used to determine characteristics associated with these combinations. RESULTS: The highest medication users during the study were for the cardiovascular (2003: 80.28%; 2017: 85.63%) and nervous (2003: 66.03%; 2017: 75.41%) systems. A 3-class latent model described medication use combinations: class 1: 'Cardiovascular & neurology anatomical group' (27.25%) included participants using medications of the cardiovascular and nervous systems in their later years; class 2: 'Multiple anatomical group' (16.49%) and class 3: 'Antiinfectives & multiple anatomical group' (56.27%). When compared to the reference class (class 1), the risk of participants being in class 3 was slightly higher than being in class 2 if they had > 4 general practitioner visits (RRR 2.37; 95% CI 2.08, 2.71), Department of Veterans Affairs' coverage (RRR 1.59; 95% CI 1.36, 1.86), ≥ 4 chronic diseases (RRR 3.16; 95% CI 2.56, 3.90) and were frail (RRR 1.47; 95% CI 1.27, 1.69). CONCLUSION: Identification of combinations of medication use may provide opportunities to develop multimorbidity guidelines and target medication reviews, and may help reduce medication load for older individuals.

A comparison of exosome purification methods using serum of Marek's disease virus (MDV)-vaccinated and -tumor-bearing chickens.

Marek's disease (MD) is an alphaherpesvirus (Marek's disease virus, MDV)-induced pathology of chickens associated with paralysis, immunosuppression, neurological signs, and T-cell lymphomas. MD is controlled in poultry production via live attenuated vaccines. The purpose of the current study was to compare methods for precipitating exosomes from vaccinated and protected chicken sera (VEX) and tumor-bearing chicken sera (TEX) for biomarker analysis of vaccine-induced protection and MD lymphomas respectively. A standard polyethylene glycol (PEG, 8%) method was compared to a commercial reagent (total exosome isolation reagent, TEI) for exosome yield and RNA content. Although exosomes purified by PEG or TEI were comparable in size and morphology, TEI-reagent yielded 3-4-fold greater concentration. Relative expression of 8 out of 10 G. gallus- and MDV1-encoded miRNAs examined displayed significant difference depending upon the precipitation method used. Standard PEG yields comparable, albeit lower amounts of exosomes than the TEI-reagent and a distinctive miRNA composition.

Comparison of exosomes purified via ultracentrifugation (UC) and Total Exosome Isolation (TEI) reagent from the serum of Marek's disease virus (MDV)-vaccinated and tumor-bearing chickens.

Extracellular vesicles (EVs) is a collective term used to refer microparticles, exosomes, and apoptotic bodies produced by a variety of cells and released into interstitial spaces and bodily fluids. Serum exosomes can serve as invaluable biomarkers, containing m/miRNAs, lipids, and proteins, indicative of various conditions. There are currently limited studies on the characterization and mutual consensus of biomarker profiles of serum exosomes purified by different methods. Here we compared the advantages and disadvantages of two commonly used serum exosome purification procedures including ultracentrifugation (UC) and Total Exosome Isolation (TEI) reagent, by analyzing exosome size distribution, concentration, morphology and miRNA expression profiles. Serum was obtained from Marek's disease virus (MDV)-infected chickens that were either vaccinated against Marek's disease (MD), and thus protected, or unvaccinated and bearing MDV-induced tumors. Nanoparticle tracking analysis (NTA) and Transmission Electron Microscopy (TEM) were performed to evaluate particle size, concentration, and morphological integrity, respectively. Our results indicate that the size distribution of particles purified by either procedure is consistent with that of exosomes (30-150 nm). TEI reagent generated higher yields and co-isolated additional EV populations that are slightly larger (∼180 nm). Based on the miRNA expression profiles from a previous high throughput sequencing experiment of exosome small RNAs, we selected six cellular and four MDV1 miRNAs, to validate their expression in UC- and TEI-purified exosomes. miRNA expression profiles displayed relative correlation between the two procedures, but distinctive differences were observed in abundance with TEI-purified exosomes showing higher miRNA expression consistent with higher yield than those purified by UC. TEI-purified exosomes from vaccinated chickens exhibited greater expression of tumor suppressor miRNA, gga-mir-146b and least expression of oncomiR, gga-mir-21 compared to those obtained from tumor-bearing chickens. We propose that gga-mir-146 and -21 can serve as serum exosome biomarkers for vaccine-induced protection and MD tumors respectively.