Isolation, Characterization, and Stability of Discretely-Sized Nanolipoprotein Particles Assembled with Apolipophorin-III

Background: Nanolipoprotein particles (NLPs) are discoidal, nanometer-sized particles comprised of self-assembled phospholipid membranes and apolipoproteins. NLPs assembled with human apolipoproteins have been used for myriad biotechnology applications, including membrane protein solubilization, drug delivery, and diagnostic imaging. To expand the repertoire of lipoproteins for these applications, insect apolipophorin-III (apoLp-III) was evaluated for the ability to form discretely-sized, homogeneous, and stable NLPs. Methodology: Four NLP populations distinct with regards to particle diameters (ranging in size from 10 nm to.25 nm) and lipid-to-apoLp-III ratios were readily isolated to high purity by size exclusion chromatography. Remodeling of the purified NLP species over time at 4uC was monitored by native gel electrophoresis, size exclusion chromatography, and atomic force microscopy. Purified 20 nm NLPs displayed no remodeling and remained stable for over 1 year. Purified NLPs with 10 nm and 15 nm diameters ultimately remodeled into 20 nm NLPs over a period of months. Intra-particle chemical cross-linking of apoLp-III stabilized NLPs of all sizes. Conclusions: ApoLp-III-based NLPs can be readily prepared, purified, characterized, and stabilized, suggesting their utilit

Prevalence of vasculare risk factors in different stages of prodromal Alzheimer’s disease and its influence on cognitive decline

peer reviewe

Architectural barriers to a safe education of persons with disabilities

Problem niepełnosprawności jest coraz bardziej widoczny w mediach oraz codziennych aspektach życia. Coraz częściej budynki użyteczności publicznej są wyposażone w pochylnie oraz podnośniki, dostosowywane do potrzeb osób niepełnosprawnych. Komunikacja miejska staję się coraz bardziej dostępna, rynek pracy jest bardziej otwarty. Pomimo znacznego postępu, niestety jednak nadal na każdym kroku istnieją bariery, które stawiają niepełnosprawność, jako poważne wyzwanie dla całego społeczeństwa. Wiele obszarów egzystencji społecznej jest nadal mało dostępna dla osób niepełnosprawnych, które pomimo barier technicznych, muszą walczyć z uprzedzeniami wobec swojej osoby. Dzieci niepełnosprawne w świetle prawa mogą uczęszczać do szkoły ogólnodostępnej, niestety w większości przypadków, szkoła taka jest dla nich niedostępna. Placówki edukacyjne w większości przypadku są budynkami zbudowanymi przed rokiem 1994, rzadko otoczenie zewnętrzne jest dostosowywane dla osób niepełnosprawnych, brak podstawowych podjazdów oraz bezpiecznych dróżek, nie wspominając o przestrzeni wewnętrznej, wszechobecne schody oraz wąskie korytarze, stają się przeszkodą nie do pokonania. Celem niniejszego artykułu jest przedstawienie rozwiązań techniczno-organizacyjnych, likwidujących bariery architektoniczne, a których wdrożenie ułatwia osobom niepełnosprawnym dostęp do budynku oraz korzystanie z usług oferowanych przez placówkę edukacyjną.The problem of disability is becoming more and more visible in the media and everyday aspects of life. Increasingly, public buildings are equipped with ramps and lifts, adapted to the needs of people with disabilities. Public transportation is becoming more available, the labor market is more open. Despite considerable progress, but still at every step there are barriers that put disability, as a serious challenge for the whole of society. Many areas of social existence is still very accessible to people with disabilities who, despite the technical barriers must fight the prejudice against his people. Children with disabilities, under the law, can attend a mainstream school, but in most cases this is the school for them unavailable. Educational in most cases are buildings built before 1994 , the external environment is rarely adapted for people with disabilities, lack of basic driveways and paths safe, not to mention the internal space, the ubiquitous stairs and narrow hallways, become an insurmountable obstacle . The purpose of this article is to provide technical and organizational solutions, eliminating architectural barriers and the implementation easier for people with disabilities access to the building and use of the services offered by the educational institution

Serum metabolites related to cardiorespiratory fitness, physical activity energy expenditure, sedentary time and vigorous activity.

The aim of our study was to investigate the relationship between objectively measured physical activity (PA) and cardiorespiratory fitness (CRF) and serum metabolites measured by targeted metabolomics in a population- based study. A total of 100 subjects provided 2 fasting blood samples and engaged in a CRF and PA measurement at 2 visits 4 months apart. CRF was estimated from a step test, whereas physical activity energy expenditure (PAEE), time spent sedentary and time spend in vigorous activity were measured by a combined heart rate and movement sensor for a total of 8 days. Serum metabolite concentrations were determined by flow injection analysis tandem mass spectrometry (FIA-MS/MS). Linear mixed models were applied with multivariable adjustment and p-values were corrected for multiple testing. Furthermore, we explored the associations between CRF, PA and two metabolite factors that have previously been linked to risk of Type 2 diabetes. CRF was associated with two phosphatidylcholine clusters independently of all other exposures. Lysophosphatidylcholine C14:0 and methionine were significantly negatively associated with PAEE and sedentary time. CRF was positively associated with the Type 2 diabetes protective factor. Vigorous activity was positively associated with the Type 2 diabetes risk factor in the mutually adjusted model. Our results suggest that CRF and PA are associated with serum metabolites, especially CRF with phosphatidylcholines and with the Type 2 diabetes protective factor. PAEE and sedentary time were associated with methionine. The identified metabolites could be potential mediators of the protective effects of CRF and PA on chronic disease risk

Crystal structure of human apolipoprotein A-I: Insights into its protective effect against cardiovascular diseases

Despite three decades of extensive studies on human apolipoprotein A-I (apoA-I), the major protein component in high-density lipoproteins, the molecular basis for its antiatherogenic function is elusive, in part because of lack of a structure of the full-length protein. We describe here the crystal structure of lipid-free apoA-I at 2.4 Å. The structure shows that apoA-I is comprised of an N-terminal four-helix bundle and two C-terminal helices. The N-terminal domain plays a prominent role in maintaining its lipid-free conformation, indicating that mutants with truncations in this region form inadequate models for explaining functional properties of apoA-I. A model for transformation of the lipid-free conformation to the high-density lipoprotein-bound form follows from an analysis of solvent-accessible hydrophobic patches on the surface of the structure and their proximity to the hydrophobic core of the four-helix bundle. The crystal structure of human apoA-I displays a hitherto-unobserved array of positively and negatively charged areas on the surface. Positioning of the charged surface patches relative to hydrophobic regions near the C terminus of the protein offers insights into its interaction with cell-surface components of the reverse cholesterol transport pathway and antiatherogenic properties of this protein. This structure provides a much-needed structural template for exploration of molecular mechanisms by which human apoA-I ameliorates atherosclerosis and inflammatory diseases

Linking diet, physical activity, cardiorespiratory fitness and obesity to serum metabolite networks: Findings from a population-based study.

Background/Objective   It is not yet resolved how lifestyle factors and intermediate phenotypes interrelate with metabolic pathways. We aimed to investigate the associations between diet, physical activity, cardiorespiratory fitness and obesity with serum metabolite networks in a population-based study. Subjects/Methods   The present study included 2380 participants of a randomly drawn subcohort of the European Prospective Investigation into Cancer and Nutrition (EPIC)-Potsdam. Targeted metabolomics was used to measure 127 serum metabolites. Additional data was available including anthropometric measurements, dietary assessment including intake of whole grain bread, coffee and cake and cookies by food frequency questionnaire, and objectively measured physical activity energy expenditure and cardiorespiratory fitness in a subsample of 100 participants. In a data-driven approach Gaussian graphical modeling was used to draw metabolite networks and depict relevant associations between exposures and serum metabolites. In addition, the relationship of different exposure-metabolite networks was estimated. Results   In the serum metabolite network, the different metabolite classes could be separated. There was a big group of phospholipids and acylcarnitines, a group of amino acids, and C6-sugar. Amino acids were particularly positively associated with cardiorespiratory fitness and physical activity. C6-sugar and acylcarnitines were positively associated with obesity and inversely with intake of whole grain bread. Phospholipids showed opposite associations with obesity and coffee intake. Metabolite networks of coffee intake and obesity were strongly inversely correlated (BMI: r=−0.57 and waist circumference: r=−0.59). A strong positive correlation was observed between metabolite networks of BMI and waist circumference (r=0.99), as well as the metabolite networks of cake and cookie intake with cardiorespiratory fitness and intake of whole grain bread (r=0.52 and r=0.50; respectively). Conclusions  Lifestyle factors and phenotypes seem to interrelate in various metabolic pathways. A possible protective effect of coffee could be mediated via counterbalance of pathways of obesity involving hepatic phospholipids. Experimental studies should validate the biological mechanism

Cross-sectional associations of objectively measured physical activity, cardiorespiratory fitness and anthropometry in european adults

Objective: To quantify the independent associations between objectively measured physical activity (PA), cardiorespiratory fitness (CRF), and anthropometry in European men and women. Methods: 2,056 volunteers from 12 centers across Europe were fitted with a heart rate and movement sensor at 2 visits 4 months apart for a total of 8 days. CRF (ml/kg/min) was estimated from an 8 minute ramped step test. A cross-sectional analysis of the independent associations between objectively measured PA (m/s2 /d), moderate and vigorous physical activity (MVPA) (%time/d), sedentary time (%time/d), CRF, and anthropometry using sex stratified multiple linear regression was performed. Results: In mutually adjusted models, CRF, PA, and MVPA were inversely associated with all anthropometric markers in women. In men, CRF, PA, and MVPA were inversely associated with BMI, whereas only CRF was significantly associated with the other anthropometric markers. Sedentary time was positively associated with all anthropometric markers, however, after adjustment for CRF significant in women only. Conclusion: CRF, PA, MVPA, and sedentary time are differently associated with anthropometric markers in men and women. CRF appears to attenuate associations between PA, MVPA, and sedentary time. These observations may have implications for prevention of obesity