| 1. |
- Barreiro, Karina, et al.
(author)
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Urinary extracellular vesicles : Assessment of pre-analytical variables and development of a quality control with focus on transcriptomic biomarker research
- 2021
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In: Journal of Extracellular Vesicles. - : Wiley. - 2001-3078. ; 10:12
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Journal article (peer-reviewed)abstract
- Urinary extracellular vesicles (uEV) are a topical source of non-invasive biomarkers for health and diseases of the urogenital system. However, several challenges have become evident in the standardization of uEV pipelines from collection of urine to biomarker analysis. Here, we studied the effect of pre-analytical variables and developed means of quality control for uEV isolates to be used in transcriptomic biomarker research. We included urine samples from healthy controls and individuals with type 1 or type 2 diabetes and normo-, micro- or macroalbuminuria and isolated uEV by ultracentrifugation. We studied the effect of storage temperature (-20°C vs. -80°C), time (up to 4 years) and storage format (urine or isolated uEV) on quality of uEV by nanoparticle tracking analysis, electron microscopy, Western blotting and qPCR. Urinary EV RNA was compared in terms of quantity, quality, and by mRNA or miRNA sequencing. To study the stability of miRNA levels in samples isolated by different methods, we created and tested a list of miRNAs commonly enriched in uEV isolates. uEV and their transcriptome were preserved in urine or as isolated uEV even after long-term storage at -80°C. However, storage at -20°C degraded particularly the GC-rich part of the transcriptome and EV protein markers. Transcriptome was preserved in RNA samples extracted with and without DNAse, but read distributions still showed some differences in e.g. intergenic and intronic reads. MiRNAs commonly enriched in uEV isolates were stable and concordant between different EV isolation methods. Analysis of never frozen uEV helped to identify surface characteristics of particles by EM. In addition to uEV, qPCR assays demonstrated that uEV isolates commonly contained polyoma viruses. Based on our results, we present recommendations how to store and handle uEV isolates for transcriptomics studies that may help to expedite standardization of the EV biomarker field.
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| 2. |
- Garthwaite, Taru, et al.
(author)
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Associations of sedentary time, physical activity, and fitness with muscle glucose uptake in adults with metabolic syndrome
- 2022
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In: Scandinavian Journal of Medicine and Science in Sports. - West Sussex : John Wiley & Sons. - 0905-7188 .- 1600-0838. ; 33:3, s. 353-358
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Journal article (peer-reviewed)abstract
- Objective: The objective of the study was to investigate the associations of sedentary time, physical activity, and cardiorespiratory fitness with skeletal muscle glucose uptake (GU). Methods: Sedentary time and physical activity were measured with accelerometers and VO2max with cycle ergometry in 44 sedentary adults with metabolic syndrome. Thigh muscle GU was determined with [18F]FDG-PET imaging. Results: Sedentary time (β = −0.374), standing (β = 0.376), steps (β = 0.351), and VO2max (β = 0.598) were associated with muscle GU when adjusted for sex, age, and accelerometer wear time. Adjustment for body fat-% turned all associations non-significant. Conclusion: Body composition is a more important determinant of muscle GU in this population than sedentary time, physical activity, or fitness. © 2022 The Authors. Scandinavian Journal of Medicine & Science In Sports published by John Wiley & Sons Ltd.
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| 3. |
- Garthwaite, Taru, et al.
(author)
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Effects of reduced sedentary time on cardiometabolic health in adults with metabolic syndrome : A three-month randomized controlled trial
- 2022
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In: Journal of Science and Medicine in Sport. - Chatswood : Elsevier Ltd. - 1440-2440 .- 1878-1861. ; 25:7, s. 579-585
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Journal article (peer-reviewed)abstract
- Objectives: To investigate if reducing sedentary behavior improves cardiometabolic biomarkers in adults with metabolic syndrome. Design: Randomized controlled trial. Methods: Sixty-four sedentary middle-aged adults with metabolic syndrome were randomized into intervention (INT; n = 33) and control (CON; n = 31) groups. INT was guided to limit sedentary behavior by 1 h/day through increased standing and light-intensity physical activity. CON was instructed to maintain usual habits. Sedentary behavior, breaks in sedentary behavior, standing, and physical activity were measured with hip-worn accelerometers for three months. Fasting blood sampling and measurements of anthropometrics, body composition, and blood pressure were performed at baseline and at three months. Linear mixed models were used for statistical analyses. Results: INT reduced sedentary behavior by 50 (95% CI: 24, 73) min/day by increasing light-intensity and moderate-to-vigorous physical activity (19 [8, 30] and 24 [14, 34] min/day, respectively). Standing increased also, but non-significantly (6 [−11, 23] min/day). CON maintained baseline activity levels. Significant intervention effects favoring INT occurred in fasting insulin (INT: 83.4 [68.7, 101.2] vs. CON: 102.0 [83.3, 125.0] pmol/l at three months), insulin resistance (HOMA-IR; 3.2 [2.6, 3.9] vs. 4.0 [3.2, 4.9]), HbA1c (37 [36, 38] vs. 38 [37, 39] mmol/mol), and liver enzyme alanine aminotransferase (28 [24, 33] vs. 33 [28, 38] U/l). Conclusions: Reducing sedentary behavior by 50 min/day and increasing light-intensity and moderate-to-vigorous activity showed benefits in several cardiometabolic biomarkers in adults with metabolic syndrome. Replacing some of the daily sedentary behavior with light-intensity and moderate-to-vigorous physical activity may help in cardiometabolic disease prevention in risk populations. © 2022 The Authors
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| 4. |
- Garthwaite, Taru, et al.
(author)
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Standing is associated with insulin sensitivity in adults with metabolic syndrome
- 2021
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In: Journal of Science and Medicine in Sport. - Chatswood : Elsevier. - 1440-2440 .- 1878-1861. ; 24:12, s. 1255-1260
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Journal article (peer-reviewed)abstract
- Objectives: To determine how components of accelerometer-measured sedentary behavior (SB) and physical activity (PA), and fitness are associated with insulin sensitivity in adults with metabolic syndrome. Design: Cross-sectional. Methods: Target population was middle-aged (40–65 years) sedentary adults with metabolic syndrome. SB, breaks in SB, standing, and PA were measured for four weeks with hip-worn accelerometers. VO2max (ml/min/kg) was measured with maximal cycle ergometry. Insulin sensitivity was determined by hyperinsulinaemic-euglycaemic clamp (M-value) and fasting blood sampling (HOMA-IR, insulin). Multivariable regression was used for analyses. Results: Sixty-four participants (37 women; 58.3 [SD 6.8] years) were included. Participants spent 10.0 (1.0) h sedentary, 1.8 (0.6) h standing, and 2.7 (0.6) h in PA and took 5149 (1825) steps and 29 (8) breaks daily. In sex-, age- and accelerometer wear time-adjusted model SB, standing, steps and VO2max were associated with M-value (β = −0.384; β = 0.400; β = 0.350; β = 0.609, respectively), HOMA-IR (β = 0.420; β = −0.548; β = −0.252; β = −0.449), and insulin (β = 0.433; β = −0.541; β = −0.252; β = −0.453); all p-values < 0.05. Breaks associated only with M-value (β = 0.277). When further adjusted for body fat %, only standing remained significantly associated with HOMA-IR (β = −0.381) and insulin (β = −0.366); significance was maintained even when further adjusted for SB, PA and fitness. Light and moderate-to-vigorous PA were not associated with insulin sensitivity. Conclusions: Standing is associated with insulin sensitivity markers. The association with HOMA-IR and insulin is independent of adiposity, PA, SB and fitness. Further studies are warranted, but these findings encourage replacing sitting with standing for potential improvements in insulin sensitivity in adults at increased type 2 diabetes risk. © 2021 The Authors.
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| 5. |
- Haapala, Eero A., et al.
(author)
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Association between cardiorespiratory fitness and metabolic health in overweight and obese adults
- 2022
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In: Journal of Sports Medicine and Physical Fitness. - Turin : Edizioni Minerva Medica. - 0022-4707 .- 1827-1928. ; 62:11, s. 1526-1533
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Journal article (peer-reviewed)abstract
- BACKGROUND: Cardiorespiratory fitness (CRF) has been inversely associated with insulin resistance and clustering of cardiometabolic risk factors among overweight and obese individuals. However, most previous studies have scaled CRF by body mass (BM) possibly inflating the association between CRF and cardiometabolic health. We investigated the associations of peak oxygen uptake (V?O2peak) and peak power output (Wpeak) scaled either by BM-1, fat free mass (FFM-1), or by allometric methods with individual cardiometabolic risk factors and clustering of cardiometabolic risk factors in 55 overweight or obese adults with metabolic syndrome. METHODS: VO2peak and Wpeak were assessed by a maximal cycle ergometer exercise test. FFM was measured by air displacement plethysmo- graph and glucose, insulin, HbA1c, triglycerides, and total, LDL, and HDL cholesterol from fasting blood samples. HOMA-IR and metabolic syndrome score (MetS) were computed. RESULTS: VO2peak and Wpeak scaled by BM-1 were inversely associated with insulin (β=-0.404 to -0.372, 95% CI: -0.704 to -0.048), HOMAIR (β=-0.442 to -0.440, 95% CI: -0.762 to -0.117), and MetS (β=-0.474 to -0.463, 95% CI: -0.798 to -0.127). Other measures of CRF were not associated with cardiometabolic risk factors. CONCLUSIONS: Our results suggest that using BM-1 as a scaling factor confounds the associations between CRF and cardiometabolic risk in overweight/obese adults with the metabolic syndrome. © 2022 EDIZIONI MINERVA MEDICA.
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| 6. |
- Laine, Saara, et al.
(author)
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Relationship between liver fat content and lifestyle factors in adults with metabolic syndrome
- 2022
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In: Scientific Reports. - London : Nature Publishing Group. - 2045-2322. ; 12:1
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Journal article (peer-reviewed)abstract
- The aim of this study was to investigate the associations between liver fat content (LFC), sedentary behaviour (SB), physical activity (PA), fitness, diet, body composition, and cardiometabolic risk factors in adults with metabolic syndrome. A total of 44 sedentary adults (mean age 58 [SD 7] years; 25 women) with overweight or obesity participated. LFC was assessed with magnetic resonance spectroscopy and imaging, SB and PA with hip-worn accelerometers (26 [SD 3] days), fitness by maximal bicycle ergometry, body composition by air displacement plethysmography and nutrient intake by 4-day food diaries. LFC was not independently associated with SB, PA or fitness. Adjusted for sex and age, LFC was associated with body fat%, body mass index, waist circumference, triglycerides, alanine aminotransferase, and with insulin resistance markers. There was and inverse association between LFC and daily protein intake, which persisted after further adjusment with body fat%. LFC is positively associated with body adiposity and cardiometabolic risk factors, and inversely with daily protein intake. SB, habitual PA or fitness are not independent modulators of LFC. However, as PA is an essential component of healthy lifestyle, it may contribute to liver health indirectly through its effects on body composition in adults with metabolic syndrome. © 2022, The Author(s).
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| 7. |
- Lener, Thomas, et al.
(author)
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Applying extracellular vesicles based therapeutics in clinical trials - an ISEV position paper.
- 2015
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In: Journal of extracellular vesicles. - : Wiley. - 2001-3078. ; 4
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Journal article (peer-reviewed)abstract
- Extracellular vesicles (EVs), such as exosomes and microvesicles, are released by different cell types and participate in physiological and pathophysiological processes. EVs mediate intercellular communication as cell-derived extracellular signalling organelles that transmit specific information from their cell of origin to their target cells. As a result of these properties, EVs of defined cell types may serve as novel tools for various therapeutic approaches, including (a) anti-tumour therapy, (b) pathogen vaccination, (c) immune-modulatory and regenerative therapies and (d) drug delivery. The translation of EVs into clinical therapies requires the categorization of EV-based therapeutics in compliance with existing regulatory frameworks. As the classification defines subsequent requirements for manufacturing, quality control and clinical investigation, it is of major importance to define whether EVs are considered the active drug components or primarily serve as drug delivery vehicles. For an effective and particularly safe translation of EV-based therapies into clinical practice, a high level of cooperation between researchers, clinicians and competent authorities is essential. In this position statement, basic and clinical scientists, as members of the International Society for Extracellular Vesicles (ISEV) and of the European Cooperation in Science and Technology (COST) program of the European Union, namely European Network on Microvesicles and Exosomes in Health and Disease (ME-HaD), summarize recent developments and the current knowledge of EV-based therapies. Aspects of safety and regulatory requirements that must be considered for pharmaceutical manufacturing and clinical application are highlighted. Production and quality control processes are discussed. Strategies to promote the therapeutic application of EVs in future clinical studies are addressed.
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| 8. |
- Maaninka, Katariina, et al.
(author)
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OxLDL sensitizes platelets for increased formation of extracellular vesicles capable of finetuning macrophage gene expression
- 2023
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In: European Journal of Cell Biology. - : Elsevier. - 0171-9335 .- 1618-1298. ; 102:2
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Journal article (peer-reviewed)abstract
- Platelet extracellular vesicles (PEVs) generated upon platelet activation may play a role in inflammatory pathologies such as atherosclerosis. Oxidized low-density lipoprotein (oxLDL), a well-known contributor to atherogenesis, activates platelets and presensitizes them for activation by other agonists. We studied the effect of oxLDL on the secretion, composition, and inflammatory functions of PEVs using contemporary EV analytics. Platelets were activated by co-stimulation with thrombin (T) and collagen (C) ± oxLDL and characterized by high-resolution flow cytometry, nanoparticle tracking analysis, proximity extension assay, western blot, and electron microscopy. The effect of PEVs on macrophage differentiation and functionality was examined by analyzing macrophage surface markers, cytokine secretion, and transcriptome. OxLDL upregulated TC-induced formation of CD61+, P-selectin+ and phosphatidylserine+ PEVs. Blocking the scavenger receptor CD36 significantly suppressed the oxLDL+TC-induced PEV formation, and HDL caused a slight but detectable suppression. The inflammatory protein cargo differed between the PEVs from stimulated and unstimulated platelets. Both oxLDL+TC- and TC-induced PEVs enhanced macrophage HLA-DR and CD86 expression and decreased CD11c expression as well as secretion of several cytokines. Pathways related to cell cycle and regulation of gene expression, and immune system signaling were overrepresented in the differentially expressed genes between TC PEV -treated vs. control macrophages and oxLDL+TC PEV -treated vs. control macrophages, respectively. In conclusion, we speculate that oxLDL and activated platelets contribute to proatherogenic processes by increasing the number of PEVs that provide an adhesive and procoagulant surface, contain inflammatory mediators, and subtly finetune the macrophage gene expression.
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| 9. |
- Norha, Jooa, et al.
(author)
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Effects of reducing sedentary behavior on cardiorespiratory fitness in adults with metabolic syndrome : A 6-month RCT
- 2023
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In: Scandinavian Journal of Medicine and Science in Sports. - Chichester : Wiley-Blackwell Publishing Inc.. - 0905-7188 .- 1600-0838. ; 33:8, s. 1452-1461
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Journal article (peer-reviewed)abstract
- Introduction:Poor cardiorespiratory fitness (CRF) is associated with adverse health outcomes. Previous observational and cross-sectional studies have suggested that reducing sedentary behavior (SB) might improve CRF. Therefore, we investigated the effects of a 6-month intervention of reducing SB on CRF in 64 sedentary inactive adults with metabolic syndrome in a non-blind randomized controlled trial.Materials and Methods:In the intervention group (INT, n = 33), the aim was to reduce SB by 1 h/day for 6 months without increasing exercise training. Control group (CON, n = 31) was instructed to maintain their habitual SB and physical activity. Maximal oxygen uptake (VO2max) was measured by maximal graded bicycle ergometer test with respiratory gas measurements. Physical activity and SB were measured during the whole intervention using accelerometers.Results:Reduction in SB did not improve VO2max statistically significantly (group × time p > 0.05). Maximal absolute power output (Wmax) did not improve significantly but increased in INT compared to CON when scaled to fat free mass (FFM) (at 6 months INT 1.54 [95% CI: 1.41, 1.67] vs. CON 1.45 [1.32, 1.59] Wmax/kgFFM, p = 0.036). Finally, the changes in daily step count correlated positively with the changes in VO2max scaled to body mass and FFM (r = 0.31 and 0.30, respectively, p < 0.05).Discussion:Reducing SB without adding exercise training does not seem to improve VO2max in adults with metabolic syndrome. However, succeeding in increasing daily step count may increase VO2max. © 2023 The Authors. Scandinavian Journal of Medicine & Science In Sports published by John Wiley & Sons Ltd.
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| 10. |
- Parkkila, Petteri, 1990, et al.
(author)
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Protein A/G-based surface plasmon resonance biosensor for regenerable antibody-mediated capture and analysis of nanoparticles
- 2022
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In: Colloids and Surfaces A: Physicochemical and Engineering Aspects. - : Elsevier BV. - 1873-4359 .- 0927-7757. ; 654
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Journal article (peer-reviewed)abstract
- Characterization of nanoparticles (NPs) and their subpopulations in heterogeneous samples is of utmost importance, for example, during the initial design of targeted NP therapies and the different phases of their production cycle. Biological NPs such as extracellular vesicles (EVs) have shown promise in improving the drug delivery capabilities compared to traditional NP-based therapies, for example, in treating cancer and neurodegenerative diseases. This work presents a general antibody-mediated surface capture and analysis protocol for NPs using a Protein A/G-functionalized surface plasmon resonance biosensor. The use of anti-streptavidin antibodies allows regenerable capture of biotin-containing NPs such as large unilamellar vesicles commonly used as drug delivery vehicles. Furthermore, the use of antibodies directed against glycophorin A and B (CD235a and b) enabled diffusion-limited specific surface capture of red blood cell-derived extracellular vesicles (RBC EVs). RBC EVs showed the efficacy of the biosensor in the determination of size and bulk concentration of NP subpopulations isolated from a complex biological matrix. The mean size of the surface-captured RBC EVs was comparable to the corresponding sizes derived for the entire EV population measured with well-established NP sizing techniques, namely, nanoparticle tracking analysis and dynamic light scattering. Taken together, the Protein A/G-functionalized biosensor provides a generic alternative to the existing NP-capturing sensors based on, for example, covalent antibody attachment, hydrophobic surfaces or biotin-capped self-assembled monolayers.
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