Additional file 1 of Association of a healthy lifestyle with mortality in older people

Additional file 1: Supplementary Table 1. Description and prevalence of lifestyle factors in the study population. Table S2. Description of the ascertainment of all-cause and cause specific mortality. Table S3. Baseline characteristics of the study participants by all-cause mortality. Table S4. Hazard ratios of all-cause mortality in relation to individual lifestyle factors (n=11,340). Table S5. Hazard ratios of CVD mortality in relation to individual lifestyle factors (n=11,340). Table S6. Hazard ratios of other mortality in relation to individual lifestyle factors (n=11,340). Table S7. Hazard ratios of cancer mortality in relation to individual lifestyle factors (n=11,340). Table S8. Hazard ratios of all-cause mortality according to combinations of healthy lifestyle factors. Table S9. Hazard ratios of all-cause mortality according to lifestyle categories within demographic, anthropometric and health subgroups. Table S10. Hazard ratios of all-cause mortality according to healthy lifestyle score (excluding former smoker and former drinker). Table S11. Description of an alternative lifestyle score containing multiple levels. Table S12. Hazard ratios of all-cause mortality according to the alternative healthy lifestyle score. Table S13. Hazard ratios of all-cause mortality according to the year-3 healthy lifestyle score

Clinical measures at baseline by age group.

ASPirin in Reducing Events in the Elderly (ASPREE), a placebo-controlled prevention trial of low dose aspirin, provided the opportunity to establish a biospecimen biobank from initially healthy persons aged 70+ years for future research. The ASPREE Healthy Ageing Biobank (ASPREE Biobank) collected, processed and stored blood and urine samples at -80degC or under nitrogen vapour at two timepoints, three years apart, from a willing subset of Australian ASPREE participants. Written informed consent included separate opt-in questions for biomarker and genetic testing. Fractionated blood and urine were aliquoted into multiple low-volume, barcoded cryotubes for frozen storage within 4 hours of collection. Specially designed and outfitted mobile laboratories provided opportunities for participation by people in regional and rural areas. Detailed, high quality demographic, physiological and clinical data were collected annually through the ASPREE trial. 12,219 participants contributed blood/urine at the first timepoint, 10,617 of these older adults provided 3-year follow-up samples, and an additional 1,712 provided saliva for DNA. The mean participant age was 74 years, 54% were female and 46% lived outside major cities. Despite geographical and logistical challenges, nearly 100% of blood/urine specimens were processed and frozen within 4 hours of collection into >1.4 million aliquots. After a median of 4.7 years, major clinical events among ASPREE Biobank participants included 332 with dementia, 613 with cardiovascular disease events, 1259 with cancer, 357 with major bleeds and 615 had died. The ASPREE Biobank houses and curates a large number of biospecimens collected prior to the clinical manifestations of major disease, and 3-year follow-up samples, all linked to high quality, extensive phenotypic information. This provides the opportunity to identify or validate diagnostic, prognostic and predictive biomarkers, and potentially study biological effectors, of ageing-related diseases or maintenance of older-age good health.</div

Physical and lifestyle measures at baseline by age group.

Physical and lifestyle measures at baseline by age group.</p

Recruitment map of ASPREE Biobank participants.

Legend: Map of Australia with rectangle of enlarged south-eastern area showing the major ASPREE Biobank recruitment sites (named) and include the number of Biobank participants recruited from each site. Processing laboratories are indicated by the dotted oval symbols, as are the regions covered by the mobile biobuses indicated by shaded oval areas. The percentages of ASPREE participants recruited to Biobank in each location, relative to the total number of ASPREE participants in each site (listed alphabetically), were Adelaide, SA (61%), Ballarat, VIC (75%), Bendigo, VIC (79%), Burnie, TAS (42%), Canberra, ACT/Southern NSW (66%), Geelong, VIC (73%), Hobart, TAS (78%), Launceston, TAS (46%), Melbourne, VIC (78%), Mildura, VIC (75%), Traralgon, VIC (77%), Warrnambool VIC/Mt Gambier, SA (83%), Wodonga/Albury, VIC/NSW (72%), Wollongong, NSW (76%). A regional site established in Shepparton, VIC closed after collecting 15 samples.</p

Biobank participant questionnaire data.

Table reports the questions (abbreviated from actual) asked of participants at the time of biospecimen collections and the numbers (and percentage of total) of participants who recorded each answer. The options were “yes” or “no” or “unsure” with the latter numbers including those questions not answered. The differences in the questions at year 3 compared with baseline collections were mainly related to use of open-label aspirin and the ASPREE clinical trial medication. (DOCX)</p

Timing of sample transport, processing & storage.

Bar graphs show elapsed time for different stages of sample preparation, categorised into 30 min or 60 min blocks. (A) Sample transport time defined as the elapsed time from sample collection to sample arrival at the processing laboratory (n = 12,219). (B) Sample processing time defined as the elapsed time from sample arrival at the laboratory to storage in a freezer (n = 12,218). (C) Total time from collection to storage in a freezer (n = 12,218). (DOCX)</p

Sample types and numbers stored at baseline and at year 3.

Sample types and numbers stored at baseline and at year 3.</p

Clinical endpoints in ASPREE Biobank participants during ASPREE (2010–2017) and ASPREE+ASPREE-XT (2010–2020).

Clinical endpoints in ASPREE Biobank participants during ASPREE (2010–2017) and ASPREE+ASPREE-XT (2010–2020).</p

Mobile laboratory used to access regional and rural/remote participants to collect and process biospecimens.

Legend: Vehicle outfitted with laboratory grade benches, refrigerated centrifuge, refrigerator, disposable equipment in secure cupboards and drawers, air conditioning, a bed readily converted to an additional bench, a chair for participants, electric step for easy access, a -80 degC portable cryoshuttle and/or LN2 resin insert cryoshipper, an external powerpoint and an external generator to provide power when not connected to main power.</p

Demographics of ASPREE Biobank participants.

ASPirin in Reducing Events in the Elderly (ASPREE), a placebo-controlled prevention trial of low dose aspirin, provided the opportunity to establish a biospecimen biobank from initially healthy persons aged 70+ years for future research. The ASPREE Healthy Ageing Biobank (ASPREE Biobank) collected, processed and stored blood and urine samples at -80degC or under nitrogen vapour at two timepoints, three years apart, from a willing subset of Australian ASPREE participants. Written informed consent included separate opt-in questions for biomarker and genetic testing. Fractionated blood and urine were aliquoted into multiple low-volume, barcoded cryotubes for frozen storage within 4 hours of collection. Specially designed and outfitted mobile laboratories provided opportunities for participation by people in regional and rural areas. Detailed, high quality demographic, physiological and clinical data were collected annually through the ASPREE trial. 12,219 participants contributed blood/urine at the first timepoint, 10,617 of these older adults provided 3-year follow-up samples, and an additional 1,712 provided saliva for DNA. The mean participant age was 74 years, 54% were female and 46% lived outside major cities. Despite geographical and logistical challenges, nearly 100% of blood/urine specimens were processed and frozen within 4 hours of collection into >1.4 million aliquots. After a median of 4.7 years, major clinical events among ASPREE Biobank participants included 332 with dementia, 613 with cardiovascular disease events, 1259 with cancer, 357 with major bleeds and 615 had died. The ASPREE Biobank houses and curates a large number of biospecimens collected prior to the clinical manifestations of major disease, and 3-year follow-up samples, all linked to high quality, extensive phenotypic information. This provides the opportunity to identify or validate diagnostic, prognostic and predictive biomarkers, and potentially study biological effectors, of ageing-related diseases or maintenance of older-age good health.</div