Aging and serum exomiR content in women-effects of estrogenic hormone replacement therapy

Exosomes participate in intercellular messaging by transporting bioactive lipid-, protein-and RNA-molecules and -complexes. The contents of the exosomes reflect the physiological status of an individual making exosomes promising targets for biomarker analyses. In the present study we extracted exosome microRNAs (exomiRs) from serum samples of premenopausal women (n = 8) and monozygotic postmenopausal twins (n = 10 female pairs), discordant for the use of estrogenic hormone replacement therapy (HRT), in order to see whether the age or/and the use of HRT associates with exomiR content. A total of 241 exomiRs were detected by next generation sequencing, 10 showing age, 14 HRT and 10 age + HRT-related differences. When comparing the groups, differentially expressed miRs were predicted to affect cell proliferation processes showing inactivation with younger age and HRT usage. MiR-106-5p, -148a-3p, -27-3p, -126-5p, -28-3p and -30a-5p were significantly associated with serum 17 beta-estradiol. MiRs formed two hierarchical clusters being indicative of positive or negative health outcomes involving associations with body composition, serum 17 beta-estradiol, fat-, glucose-and inflammatory markers. Circulating exomiR clusters, obtained by NGS, could be used as indicators of metabolic and inflammatory status affected by hormonal changes at menopause. Furthermore, the individual effects of HRT-usage could be evaluated based on the serum exomiR signature

A Genome-Wide RNAi Screen Identifies Regulators of Cholesterol-Modified Hedgehog Secretion in Drosophila

Hedgehog (Hh) proteins are secreted molecules that function as organizers in animal development. In addition to being palmitoylated, Hh is the only metazoan protein known to possess a covalently-linked cholesterol moiety. The absence of either modification severely disrupts the organization of numerous tissues during development. It is currently not known how lipid-modified Hh is secreted and released from producing cells. We have performed a genome-wide RNAi screen in Drosophila melanogaster cells to identify regulators of Hh secretion. We found that cholesterol-modified Hh secretion is strongly dependent on coat protein complex I (COPI) but not COPII vesicles, suggesting that cholesterol modification alters the movement of Hh through the early secretory pathway. We provide evidence that both proteolysis and cholesterol modification are necessary for the efficient trafficking of Hh through the ER and Golgi. Finally, we identified several putative regulators of protein secretion and demonstrate a role for some of these genes in Hh and Wingless (Wg) morphogen secretion in vivo. These data open new perspectives for studying how morphogen secretion is regulated, as well as provide insight into regulation of lipid-modified protein secretion

Early life body mass trajectories and mortality in older age: findings from the Helsinki Birth Cohort Study

Abstract Background. Overweight and obesity in childhood have been linked to an increased risk of adult mortality, but evidence is still scarce. Methods. We identified trajectories of body mass index (BMI) development in early life and investigated their mortality risk. Data come from the Helsinki Birth Cohort Study, in which 4943 individuals, born 1934-1944, had serial measures of weight and height from birth to 11 years extracted from health care records, weight and height data in adulthood, and register-based mortality data for 2000-2010. Results. Three early BMI trajectories (increasing, average, and average-to-low for men and increasing, average, and low-to-high BMI for women) were identified. Women with an increasing or low-to-high BMI (BMI lower in early childhood, later exceeded average) trajectory had an increased risk of all-cause mortality compared to those with an average BMI trajectory (HR 1.55, 95% CI 1.07-2.23; and HR 1.57, 95% CI 1.04-2.37, respectively). Similar associations were observed for cancer mortality. Among men, BMI trajectories were not associated with all-cause mortality, but those with average-to-low BMI (BMI first similar then dropped below average) had an increased risk of cancer mortality. Conclusions. An increasing BMI in early life may shorten the lifespan of maturing cohorts as they age, particularly among women

Intellectual ability in young adulthood as an antecedent of physical functioning in older age

Objectives: low cognitive ability is associated with subsequent functional disability. Whether this association extends across adult life has been little studied. The aim of this study was to examine the association between intellectual ability in young adulthood and physical functioning during a 10-year follow-up in older age.Methods: three hundred and sixty persons of the Helsinki Birth Cohort Study (HBCS) male members, born between 1934 and 1944 and residing in Finland in 1971, took part in The Finnish Defence Forces Basic Intellectual Ability Test during the first 2 weeks of their military service training between 1952 and 1972. Their physical functioning was assessed twice using the Short Form 36 (SF-36) questionnaire at average ages of 61 and 71 years. A longitudinal path model linking Intellectual Ability Test score to the physical functioning assessments was used to explore the effect of intellectual ability in young adulthood on physical functioning in older age.Results: after adjustments for age at measurement, childhood socioeconomic status and adult BMI (kg/m2), better intellectual ability total and arithmetic and verbal reasoning subtest scores in young adulthood predicted better physical functioning at age 61 years (P values <0.021). Intellectual ability total and arithmetic and verbal reasoning subtest scores in young adulthood had indirect effects on physical functioning at age 71 years (P values <0.022) through better physical functioning at age 61 years. Adjustment for main chronic diseases did not change the results materially.Conclusion: better early-life intellectual ability helps in maintaining better physical functioning in older age

The Glycosylation of the Aspartic Proteinases from Barley (Hordeum Vulgare L.) and Cardoon (Cynara Cardunculus L.)

Plant aspartic proteinases characterised at the molecular level contain one or more consensus N-glycosylation sites [Runeberg-Roos, P., Törmäkangas, K. & Östman, A. (1991) Eur. J. Biochem. 202, 102120131027; Asakura, T., Watanabe, H., Abe, K. & Arai, S. (1995) Eur. J. Biochem. 232, 77201383; Veríssimo, P., Faro, C., Moir, A. J. G., Lin, Y., Tang, J. & Pires, E. (1996) Eur. J. Biochem. 235, 76220137681. We found that the glycosylation sites are occupied for the barley (Hordeum vulgare L.) aspartic proteinase (Asn333) and the cardoon (Cynara cardunculus L.) aspartic proteinase, cardosin A (Asn70 and Asn363). The oligosaccharides from each site were released from peptide pools by enzymatic hydrolysis with peptide-N-glycanase A or by hydrazinolysis and their structures were determined by exoglycosidase sequencing combined with matrix-assisted laser desorption/ionization time of flight mass spectrometry. It was observed that 6% of the oligosaccharides from the first glycosylation site of cardosin A are of the oligomannose type. Modified type glycans with proximal Fuc and without Xyl account for about 82%, 14% and 3% of the total oligosaccharides from the first and the second glycosylation sites of cardosin A and from H. vulgare aspartic proteinase, respectively. Oligosaccharides with Xyl but without proximal Fuc were only detected in the latter proteinase (4%). Glycans with proximal Fuc and Xyl account for 6%, 86% and 92% of the total oligosaccharides from the first and second glycosylation sites of cardosin A and from H. vulgure aspartic proteinase, respectively