Publications by authors named "Suzanne Crowe"

194 Publications

Performance characteristics of an instrument-free point-of-care CD4 test (VISITECT®CD4) for use in resource-limited settings.

J Int Med Res 2020 Sep;48(9):300060520955028

YRG Centre for AIDS Research and Education, Voluntary Health Services, Chennai, India.

Objective: CD4+ T lymphocyte count remains the most common biomarker of immune status and disease progression in human immunodeficiency virus (HIV)-positive individuals. VISITECT®CD4 is an instrument-free, low-cost point-of-care CD4 test with a cut-off of 350 CD4 cells/μL. This study aimed to evaluate VISITECT®CD4 test's diagnostic accuracy.

Methods: Two hundred HIV-positive patients attending a tertiary HIV centre in South India were recruited. Patients provided venous blood for reference and VISITECT®CD4 tests. An additional finger-prick blood sample was obtained for VISITECT®CD4. VISITECT®CD4's diagnostic performance in identifying individuals with CD4 counts ≤350 cells/μL was assessed by calculating sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) taking flow cytometry as the reference.

Results: The overall agreement between VISITECT®CD4 and flow cytometry was 89.5% using venous blood and 81.5% using finger-prick blood. VISITECT®CD4 showed better performance using venous blood [sensitivity: 96.6% (95% confidence interval: 92.1%-98.9%), specificity: 70.9% (57.1%-82.4%), PPV: 89.7% (83.9%-94.0%) and NPV: 88.6% (75.4%-96.2%)] than using finger-prick blood [sensitivity: 84.8% (77.9%-90.2%), specificity: 72.7% (59.0%-83.9%), PPV: 89.1% (82.7%-93.8%) and NPV: 64.5% (51.3%-76.3%)].

Conclusion: VISITECT®CD4 performed well using venous blood, demonstrating its potential utility in decentralization of CD4 testing services in resource-constrained settings.
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http://dx.doi.org/10.1177/0300060520955028DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7536493PMC
September 2020

Phylogenetic clustering networks among heterosexual migrants with new HIV diagnoses post-migration in Australia.

PLoS One 2020 1;15(9):e0237469. Epub 2020 Sep 1.

Burnet Institute, Melbourne, Victoria, Australia.

Background: It is estimated that approximately half of new HIV diagnoses among heterosexual migrants in Victoria, Australia, were acquired post-migration. We investigated the characteristics of phylogenetic clusters in notified cases of HIV among heterosexual migrants.

Methods: Partial HIV pol sequences obtained from routine clinical genotype tests were linked to Victorian HIV notifications with the following exposures listed on the notification form: heterosexual sexual contact, injecting drug use, bisexual sexual contact, male-to male sexual contact or heterosexual sexual contact in combination with injecting drug use, unknown exposure. Those with heterosexual sexual contact as the only exposure were the focus of this study, with the other exposures included to better understand transmission networks. Additional reference sequences were extracted from the Los Alamos database. Maximum likelihood methods were used to infer the phylogeny and the robustness of the resulting tree was assessed using bootstrap analysis. Phylogenetic clusters were defined on the basis of bootstrap and genetic distance.

Results: HIV pol sequences were available for 332 of 445 HIV notifications attributed to only heterosexual sexual contact in Victoria from 2005-2014. Forty-three phylogenetic clusters containing at least one heterosexual migrant were detected, 30 (70%) of which were pairs. The characteristics of these phylogenetic clusters varied considerably by cluster size. Pairs were more likely to be composed of people living with HIV from a single country of birth (p = 0.032). Larger clusters (n≥3) were more likely to contain people born in Australian/New Zealand (p = 0.002), migrants from more than one country of birth (p = 0.013) and viral subtype-B, the most common subtype in Australia (p = 0.006). Pairs were significantly more likely to contain females (p = 0.037) and less likely to include HIV diagnoses with male-to-male sexual contact reported as a possible exposure (p<0.001) compared to larger clusters (n≥3).

Conclusion: Migrants appear to be at elevated risk of HIV acquisition, in part due to intimate relationships between migrants from the same country of origin, and in part due to risks associated with the broader Australian HIV epidemic. However, there was no evidence of large transmission clusters driven by heterosexual transmission between migrants. A multipronged approach to prevention of HIV among migrants is warranted.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0237469PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7462279PMC
October 2020

The Multiparametric Analysis of Mitochondrial Dynamics in T Cells from Cryopreserved Peripheral Blood Mononuclear Cells (PBMCs).

Methods Mol Biol 2020 ;2184:215-224

Life Sciences, Burnet Institute, Melbourne, VIC, Australia.

The analysis of mitochondrial dynamics within immune cells allows us to understand how fundamental metabolism influences immune cell functions, and how dysregulated immunometabolic processes impact biology and disease pathogenesis. For example, during infections, mitochondrial fission and fusion coincide with effector and memory T-cell differentiation, respectively, resulting in metabolic reprogramming. As frozen cells are generally not optimal for immunometabolic analyses, and given the logistic difficulties of analysis on cells within a few hours of blood collection, we have optimized and validated a simple cryopreservation protocol for peripheral blood mononuclear cells, yielding >95% cellular viability, as well as preserved metabolic and immunologic properties. Combining fluorescent dyes with cell surface antibodies, we demonstrate how to analyze mitochondrial density, membrane potential, and reactive oxygen species production in CD4 and CD8 T cells from cryopreserved clinical samples.
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http://dx.doi.org/10.1007/978-1-0716-0802-9_15DOI Listing
March 2021

Gaps in the HIV diagnosis and care cascade for migrants in Australia, 2013-2017: A cross-sectional study.

PLoS Med 2020 03 10;17(3):e1003044. Epub 2020 Mar 10.

Public Health Discipline, Burnet Institute, Melbourne, Australia.

Background: Globally, few studies compare progress toward the Joint United Nations Program on HIV/AIDS (UNAIDS) Fast-Track targets among migrant populations. Fast-Track targets are aligned to the HIV diagnosis and care cascade and entail achieving 90-90-90 (90% of people living with HIV [PLHIV] diagnosed, 90% of those diagnosed on treatment, and 90% of those on treatment with viral suppression [VS]) by 2020 and 95-95-95 by 2030. We compared cascades between migrant and nonmigrant populations in Australia.

Methods And Findings: We conducted a serial cross-sectional survey for HIV diagnosis and care cascades using modelling estimates for proportions diagnosed combined with a clinical database for proportions on treatment and VS between 2013-2017. We estimated the number of PLHIV and number diagnosed using New South Wales (NSW) and Victorian (VIC) data from the Australian National HIV Registry. Cascades were stratified by migration status, sex, HIV exposure, and eligibility for subsidised healthcare in Australia (reciprocal healthcare agreement [RHCA]). We found that in 2017, 17,760 PLHIV were estimated in NSW and VIC, and 90% of them were males. In total, 90% of estimated PLHIV were diagnosed. Of the 9,391 who were diagnosed and retained in care, most (85%; n = 8,015) were males. We excluded 38% of PLHIV with missing data for country of birth, and 41% (n = 2,408) of eligible retained PLHIV were migrants. Most migrants were from Southeast Asia (SEA; 28%), northern Europe (12%), and eastern Asia (11%). Most of the migrants and nonmigrants were males (72% and 83%, respectively). We found that among those retained in care, 90% were on antiretroviral therapy (ART), and 95% of those on ART had VS (i.e., 90-90-95). Migrants had larger gaps in their HIV diagnosis and care cascade (85-85-93) compared with nonmigrants (94-90-96). Similarly, there were larger gaps among migrants reporting male-to-male HIV exposure (84-83-93) compared with nonmigrants reporting male-to-male HIV exposure (96-92-96). Large gaps were also found among migrants from SEA (72-87-93) and sub-Saharan Africa (SSA; 89-93-91). Migrants from countries ineligible for RHCA had lower cascade estimates (83-85-92) than RHCA-eligible migrants (96-86-95). Trends in the HIV diagnosis and care cascades improved over time (2013 and 2017). However, there was no significant increase in ART coverage among migrant females (incidence rate ratio [IRR]: 1.03; 95% CI 0.99-1.08; p = 0.154), nonmigrant females (IRR: 1.01; 95% CI 0.95-1.07; p = 0.71), and migrants from SEA (IRR: 1.03; 95% CI 0.99-1.07; p = 0.06) and SSA (IRR: 1.03; 95% CI 0.99-1.08; p = 0.11). Additionally, there was no significant increase in VS among migrants reporting male-to-male HIV exposure (IRR: 1.02; 95% CI 0.99-1.04; p = 0.08). The major limitation of our study was a high proportion of individuals missing data for country of birth, thereby limiting migrant status categorisation. Additionally, we used a cross-sectional instead of a longitudinal study design to develop the cascades and used the number retained as opposed to using all individuals diagnosed to calculate the proportions on ART.

Conclusions: HIV diagnosis and care cascades improved overall between 2013 and 2017 in NSW and VIC. Cascades for migrants had larger gaps compared with nonmigrants, particularly among key migrant populations. Tracking subpopulation cascades enables gaps to be identified and addressed early to facilitate achievement of Fast-Track targets.
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http://dx.doi.org/10.1371/journal.pmed.1003044DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7064172PMC
March 2020

Trends in late and advanced HIV diagnoses among migrants in Australia; implications for progress on Fast-Track targets: A retrospective observational study.

Medicine (Baltimore) 2020 Feb;99(8):e19289

Public Health, Burnet Institute.

Achieving the Joint United Nations Program on human immunodeficiency virus (HIV)/AIDS Fast-Track targets requires additional strategies for mobile populations. We examined trends and socio-demographics of migrants (overseas-born) and Australian-born individuals presenting with late and advanced HIV diagnoses between 2008 and 2017 to help inform public health approaches for HIV testing coverage and linkage to care and treatment.We conducted a retrospective population-level observational study of individuals diagnosed with HIV in Australia and reported to the National HIV Registry. Annual proportional trends in late (CD4+ T-cell count <350 cells/μL) and advanced (CD4+ T-cell count <200 cells/μL). HIV diagnoses were determined using Poisson regression.Of 9926 new HIV diagnoses from 2008 to 2017, 84% (n = 8340) were included in analysis. Overall, 39% (n = 3267) of diagnoses were classified as late; 52% (n = 1688) of late diagnoses were advanced. Of 3317 diagnoses among migrants, 47% were late, versus 34% of Australian-born diagnoses (P < .001).The annual proportions of late (incidence rate ratio [IRR] 1.00; 95% confidence interval [CI] 0.99-1.01) and advanced HIV diagnoses (IRR 1.01; 95% CI 0.99-1.02) remained constant. Among migrants with late HIV diagnosis, the proportion reporting male-to-male sex exposure (IRR 1.05; 95% CI 1.03-1.08), non-English speaking (IRR 1.03; 95% CI 1.01-1.05), and individuals born in countries in low HIV-prevalence (IRR 1.02; 95% CI 1.00-1.04) increased. However, declines were noted among some migrants' categories such as females, heterosexual exposure, English speaking, and those born in high HIV-prevalence countries.Late HIV diagnosis remains a significant public health concern in Australia. Small declines in late diagnosis among some migrant categories are offset by increases among male-to-male exposures. Reaching the Fast-Track targets in Australia will require targeted testing and linkage to care strategies for all migrant populations, especially men who have sex with men.
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http://dx.doi.org/10.1097/MD.0000000000019289DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7034696PMC
February 2020

Modelling the impact of migrants on the success of the HIV care and treatment program in Botswana.

PLoS One 2020 15;15(1):e0226422. Epub 2020 Jan 15.

Burnet Institute, Melbourne, Australia.

Introduction: Botswana offers publicly financed HIV treatment to citizens, but not migrants, who comprised about 7% of the population in 2016. However, HIV incidence is not declining in proportion to Botswana's HIV response. In 2018, Botswana had 86% of citizens living with HIV diagnosed, 95% of people diagnosed on treatment, and 95% viral suppression among those on treatment. We hypothesised that continued exclusion of migrants is hampering reduction of HIV incidence in Botswana. Hence, we modelled the impact of including migrants in Botswana's HIV response on achieving 90-90-90 and 95-95-95 Fast-Track targets by 2020 and 2030, respectively.

Methods: The Optima HIV model, with demographic, epidemiological, and behavioural inputs, was applied to citizens of and migrants to Botswana. Projections of new HIV infections and HIV-related deaths were compared for three scenarios to the end of 2030: (1) continued status quo for HIV testing and treatment coverage, and maintenance of levels of linkage to care, loss to follow-up, and viral suppression among citizens and migrants (baseline); (2) with scaled-up budget, optimised to achieve 90-90-90 and 95-95-95 Fast-Track targets by 2020 and 2030, respectively, for citizens only; and (3) scaled-up optimised budget to achieve these targets for both citizens and migrants.

Results: A baseline of 172,000 new HIV infections and 8,400 HIV-related deaths was projected over 2020-2030. Scaling up to achieve targets among citizens only averted an estimated 48,000 infections and 1,700 deaths. Achieving targets for both citizens and migrants averted 16,000 (34%) more infections and 442 (26%) more deaths. Scaling up for both populations reduced numbers of new HIV infections and deaths by 44% and 39% respectively compared with 2010 levels. Treating migrants when scaling up in both populations was estimated to cost USD 74 million over 2020-2030.

Conclusions: Providing HIV services to migrants in Botswana could lead to further reductions in HIV incidence and deaths. However, even with an increased, optimised budget that achieves 95-95-95 targets for both citizens and migrants by 2030, the 90% incidence reduction target for 2020 will be missed. Further efficiencies and innovations will be needed to meet HIV targets in Botswana.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0226422PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6961860PMC
April 2020

Inflammatory and immunometabolic consequences of gut dysfunction in HIV: Parallels with IBD and implications for reservoir persistence and non-AIDS comorbidities.

EBioMedicine 2019 Aug 18;46:522-531. Epub 2019 Jul 18.

Life Sciences, Burnet Institute, Melbourne, Australia; School of Medical Science, RMIT University, Melbourne, Australia; Department of Microbiology and Immunology, University of Melbourne, Melbourne, Australia. Electronic address:

The gastrointestinal mucosa is critical for maintaining the integrity and functions of the gut. Disruption of this barrier is a hallmark and a risk factor for many intestinal and chronic inflammatory diseases. Inflammatory bowel disease (IBD) and HIV infection are characterized by microbial translocation and systemic inflammation. Despite the clinical overlaps between HIV and IBD, significant differences exist such as the severity of gut damage and mechanisms of immune cell homeostasis. Studies have supported the role of metabolic activation of immune cells in promoting chronic inflammation in HIV and IBD. This inflammatory response persists in HIV+ persons even after long-term virologic suppression by antiretroviral therapy (ART). Here, we review gut dysfunction and microbiota changes during HIV infection and IBD, and discuss how this may induce metabolic reprogramming of monocytes, macrophages and T cells to impact disease outcomes. Drawing from parallels with IBD, we highlight how factors such as lipopolysaccharides, residual viral replication, and extracellular vesicles activate biochemical pathways that regulate immunometabolic processes essential for HIV persistence and non-AIDS metabolic comorbidities. This review highlights new mechanisms and support for the use of immunometabolic-based therapeutics towards HIV remission/cure, and treatment of metabolic diseases.
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http://dx.doi.org/10.1016/j.ebiom.2019.07.027DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6710907PMC
August 2019

Clinical outcomes of a cohort of migrants and citizens living with human immunodeficiency virus in Botswana: implications for Joint United Nation Program on HIV and AIDS 90-90-90 targets.

Medicine (Baltimore) 2019 Jun;98(23):e15994

Independence Surgery, Gaborone, Botswana.

The aim of the study was to evaluate the human immunodeficiency virus (HIV) treatment cascade and mortality in migrants and citizens living with HIV in Botswana.Retrospective 2002 to 2016 cohort study using electronic medical records from a single center managing a high migrant case load.Records for 768 migrants and 3274 citizens living with HIV were included. Maipelo Trust, a nongovernmental organization, funded care for most migrants (70%); most citizens (85%) had personal health insurance. Seventy percent of migrants and 93% of citizens had received antiretroviral therapy (ART). At study end, 44% and 27% of migrants and citizens, respectively were retained in care at the clinic (P < .001). Among the 35% and 60% of migrants and citizens on ART respectively with viral load (VL) results in 2016, viral suppression was lower among migrants (82%) than citizens (95%) (P < .001). Citizens on ART had a median 157-unit [95% confidence interval (CI) 122-192] greater increase in CD4+ T-cell count (last minus first recorded count) than migrants after adjusting for baseline count (P < .001). Five-year survival was 92% (95% CI = 87.6-94.8) for migrants and 96% (95% CI = 95.4-97.2) for citizens. Migrants had higher mortality than citizens after entry into care (hazard ratio = 2.3, 95% CI = 1.34-3.89, P = .002) and ART initiation (hazard ratio = 2.2, 95% CI = 1.24-3.78, P = .01).Fewer migrants than citizens living with HIV in Botswana were on ART, accessed VL monitoring, achieved viral suppression, and survived. The HIV treatment cascade appears suboptimal for migrants, undermining local 90-90-90 targets. These results highlight the need to include migrants in mainstream-funded HIV treatment programs, as microepidemics can slow HIV epidemic control.
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http://dx.doi.org/10.1097/MD.0000000000015994DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6571245PMC
June 2019

People who inject drugs in Bangladesh - The untold burden!

Int J Infect Dis 2019 Jun 27;83:109-115. Epub 2019 Mar 27.

International Centre for Diarrhoeal Disease Research, Dhaka, Bangladesh; Menzies Health Institute, Gold Coast, Australia. Electronic address:

The rates of both HIV and HCV are exploding among the People Who Inject Drugs (PWID) subpopulation in the People's Republic of Bangladesh. 5,586 HIV confirmed cases have been reported since the first case of HIV was identified in 1989, of which, 865 new cases (15.5%) have been reported in the year 2017 alone. Among the new cases, 330 (38.2%) were from PWID population. The HCV prevalence is also high in Dhaka, with 40% of the PWID with unknown HIV status and 60.7% co-infected with HIV. The predominant HIV-1 strains circulating in the population are subtype C (41.4%) followed by CRF07 BC (24.2%), CRF01 AE (9.1), A1 (6.6%), and B (2.5%). HCV subtypes 3a and 3b are the most prevalent circulating strains (88.5%) among PWID. Harm reduction interventions particularly Needle Syringe Program (NSP) for PWID have been operating in Bangladesh since 1998. Opioid Substitution Therapy (OST) commenced in 2010 but only covers 2.9% of the total estimated PWID population in the country. A preliminary assessment of the needle/syringe sharing networks of HIV positive PWID was made in order to determine the HIV status among needle/syringe sharing partners. From a network of 36 HIV positive PWID seeds, 96 needle/syringe sharing partners were identified, of which 10 were HIV positive. Characterization of the nature of transmission within PWID networks is required in order to develop clinical services aimed at this vulnerable subpopulation and to halt the epidemic.
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http://dx.doi.org/10.1016/j.ijid.2019.03.009DOI Listing
June 2019

Grief reactions and coping strategies of trainee doctors working in paediatric intensive care.

Br J Anaesth 2019 07 6;123(1):74-80. Epub 2019 Mar 6.

Department of Human Factors in Patient Safety, Royal College of Surgeons of Ireland, Dublin, Ireland.

Background: The death of a child can have significant emotional effects on doctors responsible for their care. Trainee doctors working in the paediatric intensive care unit (PICU) may be particularly vulnerable. The aim of this study was to examine the emotional impact of, and grief reactions to, a child's death in PICU trainee doctors, along with coping strategies they used.

Methods: In a prospective, cross-sectional, observational study, qualitative and quantitative data were recorded on anonymised, written questionnaires. Grief severity was assessed using the Texas Revised Inventory of Grief. Emotional impact was assessed using the shortened Impact of Event Scale. The BriefCOPE tool was used to assess coping strategies. Qualitative data was analysed using conventional content analysis. Data are presented as median (inter-quartile range) or number (%).

Results: All invited trainee doctors (23 anaesthetists; 5 paediatricians) completed the questionnaire (age, 30 [29-34] yr; 13/28 [46%] female). Two (7%) doctors experienced severe grief (Texas Revised Inventory of Grief score <39), with five (18%) doctors severely affected by the deaths as measured by the Impact of Event Scale. Qualitative analysis revealed prominent themes of sadness, helplessness, guilt, shock, and concern for the bereaved family. There was limited use of coping strategies. Speaking with another trainee doctor was the principal coping strategy. Requests for debriefing sessions, greater psychological support and follow-up with the patient's family were frequently suggested.

Conclusions: Paediatric deaths evoke significant grief and emotional reactions in a subset of PICU trainee doctors. Trainee PICU doctors highlighted a lack of professional support and tailored debriefs.
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http://dx.doi.org/10.1016/j.bja.2019.01.034DOI Listing
July 2019

Qualitative assessment of South African healthcare worker perspectives on an instrument-free rapid CD4 test.

BMC Health Serv Res 2019 Feb 14;19(1):123. Epub 2019 Feb 14.

Wits Reproductive Health and HIV Institute (Wits RHI), Faculty of Health Sciences, University of the Witwatersrand, Johannesburg, South Africa.

Background: Accurate measurement of CD4 cell counts remains an important tenet of clinical care for people living with HIV. We assessed an instrument-free point-of-care CD4 test (VISITECT® CD4) based on a lateral flow principle, which gives visual results after 40 min. The test involves five steps and categorises CD4 counts as above or below 350 cells/μL. As one component of a performance evaluation of the test, this qualitative study explored the views of healthcare workers in a large women and children's hospital on the acceptability and feasibility of the test.

Methods: Perspectives on the VISITECT® CD4 test were elicited through in-depth interviews with eight healthcare workers involved in the performance evaluation at an antenatal care facility in Johannesburg, South Africa. Audio recordings were transcribed in full and analysed thematically.

Results: Healthcare providers recognised the on-going relevance of CD4 testing. All eight perceived the VISITECT® CD4 test to be predominantly user-friendly, although some felt that the need for precision and optimal concentration in performing test procedures made it more challenging to use. The greatest strength of the test was perceived to be its quick turn-around of results. There were mixed views on the semi-quantitative nature of the test results and how best to integrate this test into existing health services. Participants believed that patients in this setting would likely accept the test, given their general familiarity with other point-of-care tests.

Conclusions: Overall, the VISITECT® CD4 test was acceptable to healthcare workers and those interviewed were supportive of scale-up and implementation in other antenatal care settings. Both health workers and patients will need to be oriented to the semi-quantitative nature of the test and how to interpret the results of tests.
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http://dx.doi.org/10.1186/s12913-019-3948-xDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6376755PMC
February 2019

Performance of a Novel Low-Cost, Instrument-Free Plasma Separation Device for HIV Viral Load Quantification and Determination of Treatment Failure in People Living with HIV in Malaysia: a Diagnostic Accuracy Study.

J Clin Microbiol 2019 04 28;57(4). Epub 2019 Mar 28.

Burnet Institute, Melbourne, Australia.

HIV viral load (VL) testing is the recommended method for monitoring the response of people living with HIV and receiving antiretroviral therapy (ART). The availability of standard plasma VL testing in low- and middle-income countries (LMICs), and access to this testing, are limited by the need to use fresh plasma. Good specimen collection methods for HIV VL testing that are applicable to resource-constrained settings are needed. We assessed the diagnostic performance of the filtered dried plasma spot (FDPS), created using the newly developed, instrument-free VLPlasma device, in identifying treatment failure at a VL threshold of 1,000 copies/ml in fresh plasma. Performance was compared with that of the conventional dried blood spot (DBS). Venous blood samples from 201 people living with HIV and attending an infectious disease clinic in Malaysia were collected, and HIV VL was quantified using fresh plasma (the reference standard), FDPS, and DBS specimens. VL testing was done using the Roche Cobas AmpliPrep/Cobas TaqMan v2.0 assay. At a threshold of 1,000 copies/ml, the diagnostic performance of the FDPS was superior (sensitivity, 100% [95% confidence interval {CI}, 89.1 to 100%]; specificity, 100% [95% CI, 97.8 to 100%]) to that of the DBS (sensitivity, 100% [95% CI, 89.4 to 100%]; specificity, 36.8% [95% CI, 29.4 to 44.7%]) ( < 0.001). A stronger correlation was observed between the FDPS VL and the plasma VL ( = 0.94;  < 0.001) than between the DBS VL and the plasma VL ( = 0.85;  < 0.001). The mean difference in VL measures between the FDPS and plasma (plasma VL minus FDPS VL) was 0.127 log copies/ml (standard deviation [SD], 0.32), in contrast to -0.95 log copies/ml (SD, 0.84) between the DBS and plasma. HIV VL measurement using the FDPS outperformed that with the DBS in identifying treatment failure at a threshold of 1,000 copies/ml and compared well with the quantification of VL in plasma. The FDPS can be an attractive alternative to fresh plasma for improving access to HIV VL monitoring among people living with HIV on ART in LMICs.
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http://dx.doi.org/10.1128/JCM.01683-18DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6440787PMC
April 2019

Difficult Social Circumstances.

Authors:
Suzanne Crowe

Anesthesiology 2019 May;130(5):851-852

From the Pediatric Intensive Care Unit, Our Lady's Children's Hospital, Crumlin, Dublin, Ireland.

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http://dx.doi.org/10.1097/ALN.0000000000002592DOI Listing
May 2019

A Bioelectronic System to Measure the Glycolytic Metabolism of Activated CD4+ T Cells.

Biosensors (Basel) 2019 Jan 9;9(1). Epub 2019 Jan 9.

Life Sciences Discipline, Burnet Institute, Melbourne, VIC 3001, Australia.

The evaluation of glucose metabolic activity in immune cells is becoming an increasingly standard task in immunological research. In this study, we described a sensitive, inexpensive, and non-radioactive assay for the direct and rapid measurement of the metabolic activity of CD4+ T cells in culture. A portable, custom-built Cell Culture Metabolite Biosensor device was designed to measure the levels of acidification (a proxy for glycolysis) in cell-free CD4+ T cell culture media. In this assay, ex vivo activated CD4+ T cells were incubated in culture medium and mini electrodes were placed inside the cell free culture filtrates in 96-well plates. Using this technique, the inhibitors of glycolysis were shown to suppress acidification of the cell culture media, a response similar to that observed using a gold standard lactate assay kit. Our findings show that this innovative biosensor technology has potential for applications in metabolic research, where acquisition of sufficient cellular material for ex vivo analyses presents a substantial challenge.
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http://dx.doi.org/10.3390/bios9010010DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6468583PMC
January 2019

Field Performance and Diagnostic Accuracy of a Low-Cost Instrument-Free Point-of-Care CD4 Test (Visitect CD4) Performed by Different Health Worker Cadres among Pregnant Women.

J Clin Microbiol 2019 02 30;57(2). Epub 2019 Jan 30.

Burnet Institute, Melbourne, Victoria, Australia.

Measuring CD4 counts remains an important component of HIV care. The Visitect CD4 is the first instrument-free low-cost point-of-care CD4 test with results interpreted visually after 40 min, providing a result of ≥350 CD4 cells/mm The field performance and diagnostic accuracy of the test was assessed among HIV-infected pregnant women in South Africa. A nurse performed testing at the point-of-care using both venous and finger-prick blood, and a counselor and laboratory staff tested venous blood in the clinic laboratory (four Visitect CD4 tests/participant). Performance was compared to the mean CD4 count from duplicate flow cytometry tests on venous blood (FACSCalibur Trucount). In 2017, 156 patients were enrolled, providing a total of 624 Visitect CD4 tests (468 venous and 156 finger-prick samples). Of 624 tests, 28 (4.5%) were inconclusive. Generalized linear mixed modeling showed better performance of the test on venous blood (sensitivity = 81.7%; 95% confidence interval [CI] = 72.3 to 91.1]; specificity = 82.6%, 95% CI = 77.1 to 88.1) than on finger-prick specimens (sensitivity = 60.7%; 95% CI = 45.0 to 76.3; specificity = 89.5%, 95% CI = 83.2 to 95.8;  = 0.001). No difference in performance was detected by cadre of health worker ( = 0.113) or between point-of-care versus laboratory-based testing ( = 0.108). Adequate performance of Visitect CD4 with different operators and at the point of care, with no need of electricity or instrument, shows the potential utility of this device, especially for facilitating decentralization of CD4 testing services in rural areas.
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http://dx.doi.org/10.1128/JCM.01277-18DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6355532PMC
February 2019

Induction of HIF-1α by HIV-1 Infection in CD4 T Cells Promotes Viral Replication and Drives Extracellular Vesicle-Mediated Inflammation.

mBio 2018 09 11;9(5). Epub 2018 Sep 11.

INBIRS, Facultad de Medicina, Buenos Aires, Argentina

Chronic immune activation and inflammation are hallmarks of HIV-1 infection and a major cause of serious non-AIDS events in HIV-1-infected individuals on antiretroviral treatment (ART). Herein, we show that cytosolic double-stranded DNA (dsDNA) generated in infected CD4 T cells during the HIV-1 replication cycle promotes the mitochondrial reactive oxygen species (ROS)-dependent stabilization of the transcription factor hypoxia-inducible factor 1α (HIF-1α), which in turn, enhances viral replication. Furthermore, we show that induction of HIF-1α promotes the release of extracellular vesicles (EVs). These EVs foster inflammation by inducing the secretion of gamma interferon by bystander CD4 T cells and secretion of interleukin 6 (IL-6) and IL-1β by bystander macrophages through an HIF-1α-dependent pathway. Remarkably, EVs obtained from plasma samples from HIV-1-infected individuals also induced HIF-1α activity and inflammation. Overall, this study demonstrates that HIF-1α plays a crucial role in HIV-1 pathogenesis by promoting viral replication and the release of EVs that orchestrate lymphocyte- and macrophage-mediated inflammatory responses. Human immunodeficiency virus type 1 (HIV-1) is a very important global pathogen that preferentially targets CD4 T cells and causes acquired immunodeficiency syndrome (AIDS) if left untreated. Although antiretroviral treatment efficiently suppresses viremia, markers of immune activation and inflammation remain higher in HIV-1-infected patients than in uninfected individuals. The hypoxia-inducible factor 1α (HIF-1α) is a transcription factor that plays a fundamental role in coordinating cellular metabolism and function. Here we show that HIV-1 infection induces HIF-1α activity and that this transcription factor upholds HIV-1 replication. Moreover, we demonstrate that HIF-1α plays a key role in HIV-1-associated inflammation by promoting the release of extracellular vesicles which, in turn, trigger the secretion of inflammatory mediators by noninfected bystander lymphocytes and macrophages. In summary, we identify that the coordinated actions of HIF-1α and extracellular vesicles promote viral replication and inflammation, thus contributing to HIV-1 pathogenesis.
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http://dx.doi.org/10.1128/mBio.00757-18DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6134101PMC
September 2018

Anesthesia consensus on clinical parameters for the timing of surgical repair in congenital diaphragmatic hernia.

Paediatr Anaesth 2018 08;28(8):751-752

Department of Anesthesia & Critical Care Medicine, Our Lady's Children's Hospital, Crumlin, Dublin, Ireland.

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http://dx.doi.org/10.1111/pan.13459DOI Listing
August 2018

Polymorphism rs1385129 Within Glut1 Gene Is Linked to Poor CD4+ T Cell Recovery in Antiretroviral-Treated HIV+ Individuals.

Front Immunol 2018 17;9:900. Epub 2018 May 17.

Centre for Biomedical Research, Burnet Institute, Melbourne, VIC, Australia.

Untreated HIV infection is associated with progressive CD4+ T cell depletion, which is generally recovered with combination antiretroviral therapy (cART). However, a significant proportion of cART-treated individuals have poor CD4+ T cell reconstitution. We investigated associations between HIV disease progression and CD4+ T cell glucose transporter-1 (Glut1) expression. We also investigated the association between these variables and specific single nucleotide polymorphisms (SNPs) within the Glut1 regulatory gene AKT (rs1130214, rs2494732, rs1130233, and rs3730358) and in the Glut1-expressing gene SLC2A1 (rs1385129 and rs841853) and antisense RNA 1 region SLC2A1-AS1 (rs710218). High CD4+Glut1+ T cell percentage is associated with rapid CD4+ T cell decline in HIV-positive treatment-naïve individuals and poor T cell recovery in HIV-positive individuals on cART. Evidence suggests that poor CD4+ T cell recovery in treated HIV-positive individuals is linked to the homozygous genotype (GG) associated with SLC2A1 SNP rs1385129 when compared to those with a recessive allele (GA/AA) (odds ratio = 4.67;  = 0.04). Furthermore, poor response to therapy is less likely among Australian participants when compared against American participants (odds ratio: 0.12;  = 0.01) despite there being no difference in prevalence of a specific genotype for any of the SNPs analyzed between nationalities. Finally, CD4+Glut1+ T cell percentage is elevated among those with a homozygous dominant genotype for SNPs rs1385129 (GG) and rs710218 (AA) when compared to those with a recessive allele (GA/AA and AT/TT respectively) ( < 0.04). The heterozygous genotype associated with AKT SNP 1130214 (GT) had a higher CD4+Glut1+ T cell percentage when compared to the dominant homozygous genotype (GG) ( = 0.0068). The frequency of circulating CD4+Glut1+ T cells and the rs1385129 SLC2A1 SNP may predict the rate of HIV disease progression and CD4+ T cell recovery in untreated and treated infection, respectively.
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http://dx.doi.org/10.3389/fimmu.2018.00900DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5966582PMC
July 2019

A tale of two countries: progress towards UNAIDS 90-90-90 targets in Botswana and Australia.

J Int AIDS Soc 2018 03;21(3)

Department of Epidemiology and Preventive Medicine, Monash University, Melbourne, Vic., Australia.

UNAIDS 90-90-90 targets and Fast-Track commitments are presented as precursors to ending the AIDS epidemic by 2030, through effecting a 90% reduction in new HIV infections and AIDS-related deaths from 2010 levels (HIV epidemic control). Botswana, a low to middle-income country with the third-highest HIV prevalence, and Australia, a low-prevalence high-income country with an epidemic concentrated among men who have sex with men (MSM), have made significant strides towards achieving the UNAIDS 90-90-90 targets. These two countries provide lessons for different epidemic settings. This paper discusses the lessons that can be drawn from Botswana and Australia with respect to their success in HIV testing, treatment, viral suppression and other HIV prevention strategies for HIV epidemic control. Botswana and Australia are on target to achieving the 90-90-90 targets for HIV epidemic control, made possible by comprehensive HIV testing and treatment programmes in the two countries. As of 2015, 70% of all people assumed to be living with HIV had viral suppression in Botswana and Australia. However, HIV incidence remains above one per cent in the general population in Botswana and in MSM in Australia. The two countries have demonstrated that rapid HIV testing that is accessible and targeted at key and vulnerable populations is required in order to continue identifying new HIV infections. All citizens living with HIV in both countries are eligible for antiretroviral therapy (ART) and viral load monitoring through government-funded programmes. Notwithstanding their success in reducing HIV transmission to date, programmes in both countries must continue to be supported at current levels to maintain epidemic suppression. Scaled HIV testing, linkage to care, universal ART, monitoring patients on treatment over and above strengthened HIV prevention strategies (e.g. male circumcision and pre-exposure prophylaxis) will all continue to require funding. The progress that Botswana and Australia have made towards meeting the 90-90-90 targets is commendable. However, in order to reduce HIV incidence significantly towards 2030, there is a need for sustained HIV testing, linkage to care and high treatment coverage. Botswana and Australia provide useful lessons for developing countries with generalized epidemics and high-income countries with concentrated epidemics.
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http://dx.doi.org/10.1002/jia2.25090DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5838412PMC
March 2018

Exploiting immune cell metabolic machinery for functional HIV cure and the prevention of inflammaging.

F1000Res 2018 30;7:125. Epub 2018 Jan 30.

Centre for Biomedical Research, Macfarlane Burnet Institute for Medical Research and Public Health, Melbourne, VIC, Australia.

An emerging paradigm in immunology suggests that metabolic reprogramming and immune cell activation and functions are intricately linked. Viral infections, such as HIV infection, as well as cancer force immune cells to undergo major metabolic challenges. Cells must divert energy resources in order to mount an effective immune response. However, the fact that immune cells adopt specific metabolic programs to provide host defense against intracellular pathogens and how this metabolic shift impacts immune cell functions and the natural course of diseases have only recently been appreciated. A clearer insight into how these processes are inter-related will affect our understanding of several fundamental aspects of HIV persistence. Even in patients with long-term use of anti-retroviral therapies, HIV infection persists and continues to cause chronic immune activation and inflammation, ongoing and cumulative damage to multiple organs systems, and a reduction in life expectancy. HIV-associated fundamental changes to the metabolic machinery of the immune system can promote a state of "inflammaging", a chronic, low-grade inflammation with specific immune changes that characterize aging, and can also contribute to the persistence of HIV in its reservoirs. In this commentary, we will bring into focus evolving concepts on how HIV modulates the metabolic machinery of immune cells in order to persist in reservoirs and how metabolic reprogramming facilitates a chronic state of inflammation that underlies the development of age-related comorbidities. We will discuss how immunometabolism is facilitating the changing paradigms in HIV cure research and outline the novel therapeutic opportunities for preventing inflammaging and premature development of age-related conditions in HIV individuals.
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http://dx.doi.org/10.12688/f1000research.11881.1DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5791007PMC
January 2018

Tracheostomy as a Comfort Measure in Children With Life-Limiting Conditions.

J Palliat Care 2017 Jul/Oct;32(3-4):89-91. Epub 2017 Nov 7.

1 Paediatric Intensive Care Unit, Our Lady's Children's Hospital, Dublin, Ireland.

Palliative care for children who can expect only a short life has expanded over the last decade. Greater understanding of the measures required to ensure comfort and acceptable quality of life within the critical care environment has grown in tandem. Some more invasive interventions may be considered a "step too far" by some practitioners, including feeding gastrostomy, contracture release, and tracheostomy. Tracheostomy can facilitate a number of measures, which may enhance the brief life of the child and their family. However, tracheostomy is associated with some challenges, which may make it less suitable for some families. We discuss 3 cases where this intervention was carried out.
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http://dx.doi.org/10.1177/0825859717740064DOI Listing
November 2018

Sugar or Fat?-Metabolic Requirements for Immunity to Viral Infections.

Front Immunol 2017 16;8:1311. Epub 2017 Oct 16.

Centre for Biomedical Research, Burnet Institute, Melbourne, VIC, Australia.

The realization that an intricate link exists between the metabolic state of immune cells and the nature of the elicited immune responses has brought a dramatic evolution to the field of immunology. We will focus on how metabolic reprogramming through the use of glycolysis and fatty-acid oxidation (sugar or fat) regulates the capacity of immune cells to mount robust and effective immune responses. We will also discuss how fine-tuning sugar and fat metabolism may be exploited as a novel immunotherapeutic strategy to fight viral infections or improve vaccine efficacy.
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http://dx.doi.org/10.3389/fimmu.2017.01311DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5649203PMC
October 2017

Glut1 Expression Level on Inflammatory Monocytes is Associated With Markers of Cardiovascular Disease Risk in HIV-Infected Individuals.

J Acquir Immune Defic Syndr 2018 02;77(2):e28-e30

Centre for Biomedical Research, Macfarlane Burnet Institute for Medical Research and Public Health, Melbourne, Victoria, Australia.

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http://dx.doi.org/10.1097/QAI.0000000000001559DOI Listing
February 2018

Metabolically active CD4+ T cells expressing Glut1 and OX40 preferentially harbor HIV during in vitro infection.

FEBS Lett 2017 10 11;591(20):3319-3332. Epub 2017 Oct 11.

Centre for Biomedical Research, Burnet Institute, Melbourne, Australia.

High glucose transporter 1 (Glut1) surface expression is associated with increased glycolytic activity in activated CD4+ T cells. Phosphatidylinositide 3-kinases (PI3K) activation measured by p-Akt and OX40 is elevated in CD4+Glut1+ T cells from HIV+ subjects. TCR engagement of CD4+Glut1+ T cells from HIV+ subjects demonstrates hyperresponsive PI3K-mammalian target of rapamycin signaling. High basal Glut1 and OX40 on CD4+ T cells from combination antiretroviral therapy (cART)-treated HIV+ patients represent a sufficiently metabolically active state permissive for HIV infection in vitro without external stimuli. The majority of CD4+OX40+ T cells express Glut1, thus OX40 rather than Glut1 itself may facilitate HIV infection. Furthermore, infection of CD4+ T cells is limited by p110γ PI3K inhibition. Modulating glucose metabolism may limit cellular activation and prevent residual HIV replication in 'virologically suppressed' cART-treated HIV+ persons.
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http://dx.doi.org/10.1002/1873-3468.12843DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5658250PMC
October 2017

Assessment of metabolic and mitochondrial dynamics in CD4+ and CD8+ T cells in virologically suppressed HIV-positive individuals on combination antiretroviral therapy.

PLoS One 2017 30;12(8):e0183931. Epub 2017 Aug 30.

Centre for Biomedical Research, Burnet Institute, Melbourne, Australia.

Metabolism plays a fundamental role in supporting the growth, proliferation and effector functions of T cells. We investigated the impact of HIV infection on key processes that regulate glucose uptake and mitochondrial biogenesis in subpopulations of CD4+ and CD8+ T cells from 18 virologically-suppressed HIV-positive individuals on combination antiretroviral therapy (cART; median CD4+ cell count: 728 cells/μl) and 13 HIV seronegative controls. Mitochondrial membrane potential (MMP) and reactive oxygen species (ROS) production were also analysed in total CD4+ and CD8+ T cells. Among HIV+/cART individuals, expression of glucose transporter (Glut1) and mitochondrial density were highest within central memory and naïve CD4+ T cells, and lowest among effector memory and transitional memory T cells, with similar trends in HIV-negative controls. Compared to HIV-negative controls, there was a trend towards higher percentage of circulating CD4+Glut1+ T cells in HIV+/cART participants. There were no significant differences in mitochondrial dynamics between subject groups. Glut1 expression was positively correlated with mitochondrial density and MMP in total CD4+ T cells, while MMP was also positively correlated with ROS production in both CD4+ and CD8+ T cells. Our study characterizes specific metabolic features of CD4+ and CD8+ T cells in HIV-negative and HIV+/cART individuals and will invite future studies to explore the immunometabolic consequences of HIV infection.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0183931PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5576743PMC
October 2017

Immunometabolic and Lipidomic Markers Associated With the Frailty Index and Quality of Life in Aging HIV+ Men on Antiretroviral Therapy.

EBioMedicine 2017 Aug 18;22:112-121. Epub 2017 Jul 18.

Department of Infectious Diseases, The Alfred and Monash University, Level 2, Burnet Institute, 85 Commercial Road, Melbourne, VIC 3004, Australia; Burnet Institute, 85 Commercial Road, Melbourne, VIC 3004, Australia; Department of Microbiology and Immunology, University of Melbourne, Melbourne, Australia. Electronic address:

Chronic immune activation persists despite antiretroviral therapy (ART) in HIV+ individuals and underpins an increased risk of age-related co-morbidities. We assessed the Frailty Index in older HIV+ Australian men on ART. Immunometabolic markers on monocytes and T cells were analyzed using flow cytometry, plasma innate immune activation markers by ELISA, and lipidomic profiling by mass spectrometry. The study population consisted of 80 HIV+ men with a median age of 59 (IQR, 56-65), and most had an undetectable viral load (92%). 24% were frail, and 76% were non-frail. Frailty was associated with elevated Glucose transporter-1 (Glut1) expression on the total monocytes (p=0.04), increased plasma levels of innate immune activation marker sCD163 (OR, 4.8; CI 1.4-15.9, p=0.01), phosphatidylethanolamine PE(36:3) (OR, 5.1; CI 1.7-15.5, p=0.004) and triacylglycerol TG(16:1_18:1_18:1) (OR, 3.4; CI 1.3-9.2, p=0.02), but decreased expression of GM3 ganglioside, GM3(d18:1/18:0) (OR, 0.1; CI 0.0-0.6, p=0.01) and monohexosylceramide HexCerd(d18:1/22:0) (OR, 0.1; CI 0.0-0.5, p=0.004). There is a strong inverse correlation between quality of life and the concentration of PE(36:3) (ρ=-0.33, p=0.004) and PE(36:4) (ρ=-0.37, p=0.001). These data suggest that frailty is associated with increased innate immune activation and abnormal lipidomic profile. These markers should be investigated in larger, longitudinal studies to determine their potential as biomarkers for frailty.
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http://dx.doi.org/10.1016/j.ebiom.2017.07.015DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5552224PMC
August 2017

Frailty in men living with HIV: a cross-sectional comparison of three frailty instruments.

Antivir Ther 2018 ;23(2):117-127

Department of Infectious Diseases, The Alfred Hospital, Melbourne, Australia.

Background: Potent antiretroviral treatment has resulted in near normal life expectancy for people living with HIV. Consequently, there is an increased focus on comorbidities, frailty and quality of life.

Methods: We assessed and compared the prevalence of frailty, associated factors and relationship with quality of life in older Australian men living with HIV in a cross-sectional study using three frailty measurements. The Frailty Phenotype, Frailty Index and Edmonton Frail Scale were applied to 93 HIV-infected men aged over 50 years, on antiretroviral therapy. Multivariable ordinal logistic regression was used to analyse the associations of frailty with covariates and quality of life.

Results: The prevalence of frailty was 10.8% (n=10) using the Frailty Phenotype; 22.6% (n=21) using the Frailty Index and 15.1% (n=14) using the Edmonton Frail Scale. Frailty Phenotype-defined pre-frailty/frailty was associated with pre-1996 ART initiation (OR, 3.56; CI, 1.23, 10.36; P=0.020) and depression (OR, 3.74; CI, 1.24, 11.27; P=0.019). Osteoporosis, serious non-AIDS events and AIDS were associated with Frailty Index-defined frailty (OR, 4.84, CI, 1.27, 18.43, P=0.021; OR, 4.27, CI, 1.25, 14.58, P=0.020; OR, 4.62, CI, 1.30, 16.45, P=0.018, respectively) and Edmonton Frail Scale-defined frailty (OR, 7.51; CI, 1.55, 36.42; P=0.012; OR, 7.71; CI, 1.62, 36.75; P=0.010; OR, 8.53; CI, 1.70, 42.73; P=0.009, respectively), independent of age and current CD4 T-cell count. Frailty, defined by any of the instruments, was significantly associated with poorer quality of life (P<0.001).

Conclusions: Identifying frailty is an increasingly important contemporary consideration of HIV care related to ageing and quality of life.
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http://dx.doi.org/10.3851/IMP3185DOI Listing
September 2019

The complexities of delivering anesthesia and pediatric intensive care to a critically Ill pediatric refugee.

Paediatr Anaesth 2017 08;27(8):867-868

Department of Anesthesia, Pain Management and Pediatric Intensive Care, Our Lady's Children's Hospital, Crumlin, Dublin 12, Ireland.

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http://dx.doi.org/10.1111/pan.13155DOI Listing
August 2017