Binding of DNA origami to lipids: maximizing yield and switching via strand displacement.

Liposomes are widely used as synthetic analogues of cell membranes and for drug delivery. Lipid-binding DNA nanostructures can modify the shape, porosity and reactivity of liposomes, mediated by cholesterol modifications. DNA nanostructures can also be designed to switch conformations by DNA strand displacement. However, the optimal conditions to facilitate stable, high-yield DNA-lipid binding while allowing controlled switching by strand displacement are not known. Here, we characterized the effect of cholesterol arrangement, DNA structure, buffer and lipid composition on DNA-lipid binding and strand displacement. We observed that binding was inhibited below pH 4, and above 200 mM NaCl or 40 mM MgCl2, was independent of lipid type, and increased with membrane cholesterol content. For simple motifs, binding yield was slightly higher for double-stranded DNA than single-stranded DNA. For larger DNA origami tiles, four to eight cholesterol modifications were optimal, while edge positions and longer spacers increased yield of lipid binding. Strand displacement achieved controlled removal of DNA tiles from membranes, but was inhibited by overhang domains, which are used to prevent cholesterol aggregation. These findings provide design guidelines for integrating strand displacement switching with lipid-binding DNA nanostructures. This paves the way for achieving dynamic control of membrane morphology, enabling broader applications in nanomedicine and biophysics.

Fracture Risk and Use of Angiotensin-Converting Enzyme Inhibitors or Angiotensin II Receptor Blockers.

Medications used to treat hypertension may affect fracture risk. This study investigated fracture risk for users of angiotensin converting enzyme inhibitors (ACEI) or angiotensin II receptor blockers (ARB). Participants (899 men, median age 70.3 yr (59.9-79.1), range 50.0-96.6 yr; 574 women, median age 65.5 yr (58.1-75.4), range 50.1-94.6 yr) were from the Geelong Osteoporosis Study. Medication use was self-reported and incident fractures were ascertained using radiological reports. Bone mineral density (BMD) was measured at the femoral neck. Participants were divided into four groups: (1) non-users without hypertension, (2) non-users with hypertension, (3) ACEI users and (4) ARB users. Dosage was calculated using the defined daily dose (DDD) criteria. Participants were followed from date of visit to first fracture, death or 31 December 2016, whichever occurred first. Cox proportional hazards models were used for analyses. At least one incident fracture was sustained by 156 men and 135 women over a median(IQR) of 11.5(6.2-13.2) and 10.9(6.3-11.6) years of follow-up, respectively. In unadjusted analyses, compared to non-users without hypertension, men in all three other groups had a higher risk of fracture (Hazard Ratio (HR, 95%CI) 1.54, 1.00-2.37; 1.90, 1.18-3.05; 2.15, 1.26-3.66), for non-users with hypertension, ACEI and ARB users, respectively). Following adjustment for age, prior fracture and BMD, these associations became non-significant. A dose effect for ARB use was observed; men using lower doses had a higher risk of fracture than non-users without hypertension, in both unadjusted (2.66, 1.34-5.29) and adjusted (2.03, 1.01-4.08) analyses, but this association was not observed at higher doses. For women, unadjusted analyses showed a higher risk for ACEI users compared to non-users without hypertension (1.74, 1.07-2.83). This was explained after adjustment for age, alcohol consumption, prior fracture and BMD (1.28, 0.74-2.22). No other differences were observed. In men, lower dose (0 < DDD ≤ 1) ARB use was associated with an increased risk of fracture. ACEI or ARB use was not associated with increased risk of incident fracture in women. These findings may be important for antihypertensive treatment decisions in individuals with a high risk of fracture.

The Predictability of Frailty Associated with Musculoskeletal Deficits: A Longitudinal Study.

We investigated and quantified the predictability of frailty associated with musculoskeletal parameters. This longitudinal study included 287 men aged ≥ 50 yr at baseline (2001-2006) from the Geelong Osteoporosis Study. Baseline musculoskeletal measures included femoral neck bone mineral density (BMD), appendicular lean mass index (ALMI, kg/m2) and whole-body fat mass index (FMI, kg/m2) and lower-limb strength. Frailty at the 15 yr-follow-up (2016-2019) was defined as ≥ 3 and non-frail as < 3, of the following: unintentional weight loss, weakness, low physical activity, exhaustion, and slowness. Binary regression models and AUROC curves quantified the attributable risk of musculoskeletal factors to frailty and their predictive ability. Potential confounders included anthropometry, smoking, alcohol, FMI, socioeconomic status and comorbidities. Forty-eight (16.7%) men were frail at 15 yr-follow-up. Musculoskeletal models were better predictors of frailty compared to the referent (confounders only) model (AUROC for musculoskeletal factors 0.74 vs 0.67 for the referent model). The model with the highest AUROC (0.74; 95% CI 0.66-0.82) included BMD, ALMI and muscle strength (hip abductors) and was better than the referent model that included only lifestyle factors (p = 0.046). Musculoskeletal parameters improved the predictability model as measured by AUROC for frailty after 15 years. In general, muscle models performed better compared to bone models. Musculoskeletal parameters improved the predictability of frailty of the referent model that included lifestyle factors. Muscle deficits accounted for a greater proportion of the risk for frailty than did bone deficits. Targeting musculoskeletal health could be a possible avenue of intervention in regards to frailty.

The contribution of musculoskeletal factors to physical frailty: a cross-sectional study.

BACKGROUND: Musculoskeletal conditions and physical frailty have overlapping constructs. We aimed to quantify individual contributions of musculoskeletal factors to frailty. METHODS: Participants included 347 men and 360 women aged ≥60 yr (median ages; 70.8 (66.1-78.6) and 71.0 (65.2-77.5), respectively) from the Geelong Osteoporosis Study. Frailty was defined as ≥3, pre-frail 1-2, and robust 0, of the following; unintentional weight loss, weakness, low physical activity, exhaustion, and slowness. Measures were made of femoral neck BMD, appendicular lean mass index (ALMI, kg/m2) and whole-body fat mass index (FMI, kg/m2) by DXA (Lunar), SOS, BUA and SI at the calcaneus (Lunar Achilles Insight) and handgrip strength by dynamometers. Binary and ordinal logistic regression models and AUROC curves were used to quantify the contribution of musculoskeletal parameters to frailty. Potential confounders included anthropometry, smoking, alcohol, prior fracture, FMI, SES and comorbidities. RESULTS: Overall, 54(15.6%) men and 62(17.2%) women were frail. In adjusted-binary logistic models, SI, ALMI and HGS were associated with frailty in men (OR = 0.73, 95%CI 0.53-1.01; OR=0.48, 0.34-0.68; and OR = 0.11, 0.06-0.22; respectively). Muscle measures (ALMI and HGS) contributed more to this association than did bone (SI) (AUROCs 0.77, 0.85 vs 0.71, respectively). In women, only HGS was associated with frailty in adjusted models (OR = 0.30 95%CI 0.20-0.45, AUROC = 0.83). In adjusted ordinal models, similar results were observed in men; for women, HGS and ALMI were associated with frailty (ordered OR = 0.30 95%CI 0.20-0.45; OR = 0.56, 0.40-0.80, respectively). CONCLUSION: Muscle deficits appeared to contribute more than bone deficits to frailty. This may have implications for identifying potential musculoskeletal targets for preventing or managing the progression of frailty.

Maternal Cigarette Smoking and Congenital Upper and Lower Limb Differences: A Systematic Review and Meta-Analysis.

Maternal smoking during pregnancy has been associated with adverse effects on foetal development, including congenital limb anomalies. This systematic review aimed to provide an updated assessment of the association between maternal smoking during pregnancy and the risk of congenital limb anomalies. A systematic search was conducted to identify relevant studies published up to February 2023. Studies reporting on the relationship between maternal smoking during pregnancy and congenital digital anomalies or congenital limb reduction defects were included. Two independent reviewers screened the studies, extracted data, and assessed the quality of the included studies. Meta-analyses were performed to estimate the pooled odds ratios with 95% confidence intervals using fixed and random-effects models. In total, 37 publications comprising 11 cohort and 26 case-control studies were included in the systematic review. The meta-analysis demonstrated a significant increased risk of congenital limb reduction defects (pooled OR: 1.27, 95% CI: 1.18-1.38) in infants born to mothers who smoked during pregnancy. Similarly, a significant relationship was observed for the development of polydactyly/syndactyly/adactyly when considered as a single group (pooled OR: 1.32, 95% CI: 1.25-1.40). Yet, in contrast, no significant association was observed when polydactyly (pooled OR: 1.06, 95% CI: 0.88-1.27) or syndactyly (pooled OR: 0.91, 95% CI: 0.77-1.08) were considered individually. This systematic review provides updated evidence of a significant relationship between maternal smoking during pregnancy and increased risk of congenital limb anomalies. These findings highlight the potential detrimental effects of smoking on foetal limb development and underscore the importance of smoking cessation interventions for pregnant women to mitigate these risks.

Fluorescence Approaches for Characterizing Ion Channels in Synthetic Bilayers.

Ion channels are membrane proteins that play important roles in a wide range of fundamental cellular processes. Studying membrane proteins at a molecular level becomes challenging in complex cellular environments. Instead, many studies focus on the isolation and reconstitution of the membrane proteins into model lipid membranes. Such simpler, in vitro, systems offer the advantage of control over the membrane and protein composition and the lipid environment. Rhodopsin and rhodopsin-like ion channels are widely studied due to their light-interacting properties and are a natural candidate for investigation with fluorescence methods. Here we review techniques for synthesizing liposomes and for reconstituting membrane proteins into lipid bilayers. We then summarize fluorescence assays which can be used to verify the functionality of reconstituted membrane proteins in synthetic liposomes.

Impact microindentation in men with impaired fasting glucose and type 2 diabetes.

BACKGROUND: Individuals with type 2 diabetes (T2DM) are at increased fracture risk, with bone mineral density (BMD) measurements underestimating risk. Impact microindentation (IMI), a technique that measures bone microindentation distances, expressed as bone material strength index (BMSi), may improve fracture risk estimation in individuals with T2DM. This study describes the relationship between BMSi and glycaemia status in men and makes a comparison with bone measures from dual energy X-ray absorptiometry (DXA). MATERIAL AND METHODS: Participants were 340 men aged 33-96 yr from the Geelong Osteoporosis Study. Impaired fasting glucose (IFG) was defined using fasting plasma glucose (FPG) between 5.5 and 6.9 mmol/L. Diabetes was defined as FPG ≥ 7.0 mmol/L, use of antihyperglycemic medication and/or self-report. Two participants with type 1 diabetes were excluded. BMSi was measured using an OsteoProbe. Femoral neck (FNBMD) and lumbar spine (LSBMD) were measured using DXA (Lunar Prodigy) and trabecular bone score (TBS) was calculated (TBS iNsight Version 2.2). Using linear regression techniques, the relationship between glycaemia status and BMSi was evaluated, adjusting for other potential confounders (including lifestyle factors, clinical measurements and FNBMD). Glycaemia status was also considered as a binary variable (T2DM vs normoglycaemia and IFG). RESULTS: There were 234 (68.8%) men with normoglycaemia, 59 (17.4%) with IFG and 47 (13.8%) with diabetes. When considering glycaemia status as a binary variable, men with T2DM had lower mean BMSi compared to those without T2DM (normoglycaemia and IFG combined) (79.8; 95%CI 77.0-82.6 vs 83.0; 82.2-83.8 p = 0.043) and this difference in BMSi was independent of FNBMD. No differences were observed for either FNBMD or LSBMD; however, TBS was lower (1.177; 1.121-1.233 vs 1.256; 1.240-1.272, p = 0.015, independent of FNBMD). For glycaemia status considered in three groups, there were no differences in mean BMSi values between men with normoglycaemia, IFG and T2DM (82.9 (95%CI 82.0-83.8), 83.5 (81.8-85.2) and 79.8 (77.0-82.6), respectively; ANCOVA, p = 0.104). CONCLUSIONS: Measures reflecting bone material properties and microarchitecture (BMSi and TBS) might be better than measures of bone mass (BMD) in identifying individuals with T2DM at risk of fracture.

Association between bone measures and use of angiotensin-converting enzyme inhibitors or angiotensin II receptor blockers.

Angiotensin-converting enzyme inhibitor use in women was associated with lower femoral neck and lumbar spine bone mineral density as well as trabecular bone score compared to non-users. No differences were identified for men or for those who used ARB medications. PURPOSE: Many individuals at high fracture risk use medications such as angiotensin-converting enzyme inhibitors (ACEI) or angiotensin II receptor blockers (ARB) that could affect bone; thus, this study aimed to investigate whether there are any differences in bone mineral density (BMD) and trabecular bone score (TBS) between ACEI users, ARB users, and non-users. METHODS: Participants (685 men, 573 women) were from the Geelong Osteoporosis Study. Current medication use was self-reported. BMD at the femoral neck (FNBMD) and lumbar spine (LSBMD) were measured using DXA. TBS was calculated using TBS iNsight software. Linear regression models were used to investigate associations between ACEI or ARB use and bone measures, adjusting for other potential confounders. Due to interaction terms, data were stratified by age. RESULTS: There were 88 (12.8%) men and 41 (7.2%) women taking an ACEI medication, and 71 (10.4%) men and 76 (13.3%) women taking an ARB medication. Compared to non-users, ACEI use was associated with lower FNBMD (- 7.2%), LSBMD (- 12.2%), and TBS (- 9.0%) for women aged < 65 years. Lower TBS was also observed for women aged ≥ 65 years (- 17.3%). No differences were identified for ARB use. CONCLUSIONS: Women who used an ACEI medication had lower values for FNBMD, LSBMD and TBS compared to non-users. No differences were identified for men or for those who used ARB medications.

The Added Burden of Personality Disorder on Subsidized Australian Health Service Utilization Among Women With Mental State Disorder.

This study aimed to investigate health service utilization among women with mental state disorder only (MSD-PD), mental state disorder plus personality disorder (MSD+PD), and controls in a population-based sample. Women (n = 635) from the Geelong Osteoporosis Study completed mental health assessments and were categorized into groups (MSD-PD, MSD+PD, controls). General practitioner (mental and non-mental health encounters) and specialized mental health service utilization was ascertained from data linkage to the Medicare Benefits Schedule, Australia (01/09/2008-31/12/2012). Negative binomial and binary logistic regression models were employed to assess health service utilization differences between groups. Results indicated that women with MSD+PD had more encounters of non-mental health service utilization than women with MSD-PD and controls. Age significantly modified these relationships: women with MSD+PD and MSD-PD had more encounters of health service utilization at midlife and in the seventh decade of life. No significant differences were found in the frequency of general practitioner mental health service utilization or specialized mental health service utilization between groups. These data suggest that the presence of co-occurring PD is associated with increased health service utilization among women with other common mental health problems. Healthcare providers should be vigilant to the presence of PD when establishing management plans with patients presenting with common mental health problems.

Bone material strength index is associated with prior fracture in men with and without moderate chronic kidney disease.

BACKGROUND: Patients with chronic kidney disease (CKD) are at high risk for fracture. The ability of bone mineral density (BMD) to predict fractures in CKD patients has been inconsistent. Other measures such as trabecular bone score (TBS) and impact microindentation (IMI) may be more useful in this group. This study aimed to determine if TBS or IMI values differed between men with and without CKD and examine associations between prior fracture, TBS and IMI values. METHODS: Men (n = 343, age 33-96 yr) from the Geelong Osteoporosis Study were included. Femoral neck (FNBMD) and lumbar spine BMD (LSBMD) were measured using DXA (Lunar ProdigyPro). TBS was determined from lumbar spine scans (TBS iNsight software Version 2.2). IMI values (bone material strength index; BMSi) were measured using an OsteoProbe. CKD was defined as an eGFR<60 mL/min/1.73m2 (n = 53). Prior low trauma fractures (n = 37) were ascertained from radiological reports. Associations were examined using binary logistic regression, adjusting for potential confounders. Interaction terms were tested in all models. RESULTS: Men with CKD tended to have a higher likelihood of prior fracture (adjusted OR 2.27, 95%CI 1.02-5.01). Higher BMSi was associated with a lower likelihood of prior fracture (adjusted OR for 1SD increase: 0.70; 95%CI 0.51-0.97). This association was sustained after adjustment for FNBMD (OR 0.68; 95%CI 0.49-0.96) or LSBMD (OR 0.69; 95%CI 0.49-0.95). No interaction was detected between BMSi and CKD (p = 0.898). No associations were detected between FNBMD, LSBMD or TBS and prior fracture in either population and there were no interactions with CKD for FNBMD, LSBMD or TBS. CONCLUSIONS: BMSi was associated with prior fracture in men with and without CKD, however, FNBMD, LSBMD and TBS were not. Lack of an interaction term suggests that BMSi performed similarly in identifying the likelihood of prior fracture, regardless of CKD status. IMI may have clinical utility for assessing fracture risk in patients with CKD.