Automated image analysis detects aging in clinical-grade mesenchymal stromal cell cultures.

BACKGROUND: Senescent cells are undesirable in cell therapy products due to reduced therapeutic activity and risk of aberrant cellular effects, and methods for assessing senescence are needed. Early-passage mesenchymal stromal cells (MSCs) are known to be small and spindle-shaped but become enlarged upon cell aging. Indeed, cell morphology is routinely evaluated during MSC production using subjective methods. We have therefore explored the possibility of utilizing automated imaging-based analysis of cell morphology in clinical cell manufacturing. METHODS: An imaging system was adopted for analyzing changes in cell morphology of bone marrow-derived MSCs during long-term culture. Cells taken from the cultures at the desired passages were plated at low density for imaging, representing morphological changes observed in the clinical-grade cultures. The manifestations of aging and onset of senescence were monitored by population doubling numbers, expression of p16INK4a and p21Cip1/Waf1, ß-galactosidase activity, and telomeric terminal restriction fragment analysis. RESULTS: Cell area was the most statistically significant and practical parameter for describing morphological changes, correlating with biochemical senescence markers. MSCs from passages 1 (p1) and 3 (p3) were remarkably uniform in size, with cell areas between 1800 and 2500 µm2. At p5 the cells began to enlarge resulting in a 4.8-fold increase at p6-9 as compared to p1. The expression of p16INK4a and activity of ß-galactosidase had a strong correlation with the increase in cell area, whereas the expression of p21Cip1/Waf1 reached its maximum at the onset of growth arrest and subsequently decreased. Mean telomere length shortened at an apparently constant rate during culture, from 8.2 ± 0.3 kbp at p1, reaching 6.08 ± 0.6 kbp at senescence. CONCLUSIONS: Imaging analysis of cell morphology is a useful tool for evaluating aging in cell cultures throughout the lifespan of MSCs. Our findings suggest that imaging analysis can reproducibly detect aging-related changes in cell morphology in MSC cultures. These findings suggest that cell morphology is still a supreme measure of cell quality and may be utilized to develop new noninvasive imaging-based methods to screen and quantitate aging in clinical-grade cell cultures.

Correlates of low physical activity levels in aging men and women: the DR's EXTRA Study (ISRCTN45977199).

Recognizing correlates of low physical activity (PA) can help in targeting PA interventions for individuals who would benefit most from increasing their PA. We studied the associations of demographic, social, health, and lifestyle factors with low PA by sex in a population sample of 1,303 Finnish individuals aged 57-78 years. We defined low PA as no moderate or vigorous leisure-time PA reported in an interview. Altogether, 39% of men and 48% of women reported low PA. Satisfactory or poor perceived health and high BMI were independently associated with low PA in both sexes. In men, factors such as age, being divorced or separated, still working, having a weak social network, poor diet, and a health professional's suggestion to increase PA were associated with low PA. In women, cardiovascular disease and depressive symptoms were associated with low PA. These results can be applied in targeting PA interventions.

The current standard measure of cardiorespiratory fitness introduces confounding by body mass: the DR's EXTRA study.

OBJECTIVE: Cardiorespiratory fitness is currently estimated by dividing maximal oxygen consumption (VO(2max)) by body weight (per-weight standard). However, the statistically correct way to neutralize the effect of weight on VO(2max) in a given population is adjustment for body weight by regression techniques (adjusted standard). Our objective is to quantify the bias introduced by the per-weight standard in a population distributed across different categories of body mass. DESIGN: This is a cross-sectional study. SUBJECTS AND METHODS: Baseline measures from participants of the Dose-Responses to Exercise Training Study (DR's EXTRA), 635 men (body mass index (BMI): 19-47 kg m⁻²) and 638 women (BMI: 16-49 kg m⁻²) aged 57-78 years who performed oral glucose tolerance tests and maximal exercise stress tests with direct measurement of VO(2max). We compare the increase in VO(2max) implied by the per-weight standard with the real increase of VO(2max) per kg body weight. A linear logistic regression model estimates odds for abnormal glucose metabolism (either impaired fasting glycemia or impaired glucose tolerance or Type 2 diabetes) of the least-fit versus most-fit quartile according to both per-weight standard and adjusted standard. RESULTS: The per-weight standard implies an increase of VO(2max) with 20.9 ml min⁻¹ in women and 26.4 ml min⁻¹ in men per additional kg body weight. The true increase per kg is only 7.0 ml min⁻¹ (95% confidence interval: 5.3-8.8) and 8.0 ml min⁻¹ (95% confidence interval: 5.3-10.7), respectively. Risk for abnormal glucose metabolism in the least-fit quartile of the population is overestimated by 52% if the per-weight standard is used. CONCLUSIONS: In comparisons across different categories of body mass, the per-weight standard systematically underestimates cardiorespiratory fitness in obese subjects. Use of the per-weight standard markedly inflates associations between poor fitness and co-morbidities of obesity.

Diet, fitness and metabolic syndrome--the DR's EXTRA study.

BACKGROUND AND AIMS: To study the independent and combined associations of diet and cardiorespiratory fitness with the prevalence of the metabolic syndrome (MetS). METHODS AND RESULTS: We studied a population-based random sample of 663 men and 671 women 57-78 years of age at baseline of an ongoing randomised controlled trial. Based on a 4-day food record a diet score was created according to goals achieved (vegetables ≥400 g/day, fish ≥2 servings/week, fibre ≥14 g/1000 kcal, saturated fat <10 E%/day). Cardiorespiratory fitness was measured as maximal oxygen uptake (VO(2 max)) in a maximal symptom-limited bicycle ergometer test. MetS was defined by the National Cholesterol Education Program criteria. The lowest prevalence of MetS (5%) was observed among individuals in the highest VO(2 max) tertile and achieving 3-4 dietary goals. The highest prevalence (55%) was observed among those in the lowest VO(2 max) tertile and achieving none of the dietary goals. Among individuals in the highest VO(2 max) tertile, the odds ratio of having MetS was 0.04 (95% CI 0.02-0.10) for those achieving 3-4 dietary goals, 0.07 (0.04-0.14) for those achieving 1-2 dietary goals, and 0.16 (0.07-0.37) for those achieving none of the dietary goals compared with individuals in the lowest VO(2 max) tertile and achieving none of the goals after adjustment for confounding factors. CONCLUSION: Healthy diet and higher levels of cardiorespiratory fitness are associated with a reduced risk of having MetS. However, fitness seems to have a stronger association with MetS than diet.

Cardiorespiratory fitness in aging men and women: the DR's EXTRA study.

The aim of the study was to describe the levels and to create reference values of cardiorespiratory fitness, expressed as maximal oxygen consumption (VO(2max) ), maximal metabolic equivalents (METs) and maximal workload in aging men and women. We measured VO(2max) directly by a breath-by-breath method during a maximal exercise stress test on a bicycle ergometer with a linear workload increase of 20 W/min in a representative population sample of 672 men and 677 women aged 57-78 years. We presented the age and sex-specific categories of cardiorespiratory fitness (very low, low, medium, high and very high) based on variable distribution and non-linear regression models of VO(2max) , maximal METs and maximal workload. The linear age-related decrement of VO(2max) was -0.047 L/min/year (-2.3%) and -0.404 mL/kg/min/year (-1.6%) in men and -0.027 L/min/year (-1.9%) and -0.328 mL/kg/min/year (-1.6%) in women. After exclusion of diseased individuals, the rate of VO(2max) decrement remained similar. The number of chronic diseases (0, 1, 2 or ≥3) was inversely associated with VO(2max) in men (P<0.001) and women (P<0.001). The present study provides clinically useful reference values of cardiorespiratory fitness for primary and secondary prevention purposes in aging people.

Food consumption, nutrient intake and the risk of having metabolic syndrome: the DR's EXTRA Study.

BACKGROUND: The associations of different components of diet with metabolic syndrome (MetS) are largely unknown. We therefore studied the associations of intakes of selected food items and nutrients with the risk of having MetS. METHODS: The participants were a representative population sample of 1334 individuals (671 women, 663 men) 57-78 years of age. Dietary intake was assessed by a 4-day food record. MetS was defined by the National Cholesterol Education Program criteria. RESULTS: Consumption of vegetables, non-root vegetables, legumes and nuts berries and fish had an inverse and consumption of sausage had a direct association with the risk of having MetS in men after adjustment for age, smoking and alcohol consumption. Consumption of vegetables and non-root vegetables had an inverse and consumption of sausage had a direct association with the risk of having MetS in women after these adjustments. However, after further adjustment for maximal oxygen uptake (VO2(max)) most of these associations vanished. Men in the highest third of consumption of berries, fish, and legumes and nuts had 49, 37 and 44% lower risk of having MetS, respectively, than those in the lowest third after further adjustment for VO2(max). Women in the highest third of sausage consumption had a 72% higher risk of having MetS than non-consumers. CONCLUSIONS: Consumption of legumes and nuts, berries and fish was inversely associated with MetS in men. Consumption of sausage was directly associated with MetS in women. VO2(max) seems to be a strong confounding factor between food consumption and MetS.

Association of waist and hip circumference with 12-year progression of carotid intima-media thickness in elderly women.

OBJECTIVE: To test the hypothesis that waist and hip circumferences together provide additional information on the progression of preclinical atherosclerosis beyond either of them alone in elderly women. DESIGN: A 12-year follow-up study. SUBJECTS: A population-based sample of 102 women 60-70 years of age at baseline. MEASUREMENTS: Waist and hip circumferences and body mass index (BMI) were assessed and carotid intima-media thickness (IMT) was quantified noninvasively by ultrasonography. RESULTS: There was a significant increase in the 12-year carotid IMT progression across the thirds of waist circumference (0.080, 0.277, 0.279 mm, P=0.02 for difference) and hip circumference (0.030, 0.342, 0.260 mm, P=0.001 for difference) adjusted for conventional risk factors (age, smoking, systolic blood pressure, serum low-density lipoprotein and high-density lipoprotein cholesterol, blood glucose). The association of waist circumference with IMT progression was not significant after further adjustment for hip circumference or BMI. Further adjustment for waist circumference and BMI did not change the association of hip circumference with IMT progression. The IMT progression was greatest in women with waist circumference >83 cm and hip circumference

Comparing EST-based genetic maps between Pinus sylvestris and Pinus taeda.

A genetic map of Pinus sylvestris was constructed using ESTP (expressed sequence tag polymorphism) markers and other gene-based markers, AFLP markers and microsatellites. Part of the ESTP markers (40) were developed and mapped earlier in Pinus taeda, and additional markers were generated based on P. sylvestris sequences or sequences from other pine species. The mapping in P. sylvestris was based on 94 F(1) progeny from a cross between plus-tree parents E635C and E1101. AFLP framework maps for the parent trees were first constructed. The ESTP and other gene sequence-based markers were added to the framework maps, as well as five published microsatellite loci. The separate maps were then integrated with the aid of AFLPs segregating in both trees (dominant segregation ratios 3:1) as well as gene markers and microsatellites segregating in both parent trees (segregation ratios 1:1:1:1 or 1:2:1). The integrated map consisted of 12 groups corresponding to the P. taeda linkage groups, and additionally three and six smaller groups for E1101 and E635C, respectively. The number of framework AFLP markers in the integrated map is altogether 194 and the number of gene markers 61. The total length of the integrated map was 1,314 cM. The set of markers developed for P. sylvestris was also added to existing maps of two P. taeda pedigrees. Starting with a mapped marker from one pedigree in the source species resulted in a mapped marker in a pedigree of the other species in more than 40% of the cases, with about equal success in both directions. The maps of the two species are largely colinear, even if the species have diverged more than 70 MYA. Most cases of different locations were probably due to problems in identifying the orthologous members of gene families. These data provide a first ESTP-containing map of P. sylvestris, which can also be used for comparing this species to additional species mapped with the same markers.

Occupational health nursing in Finland.

Occupational health nursing in Finland is based on activities stated by the Occupational Health Care Act. Moreover, the occupational health nurse provides services that are based on agreements between the employer and the occupational health center. Occupational health nursing emphasizes preventive measures, such as the clarification of health hazards and job related risk factors, the arrangement of health examinations, health education, and organization of first aid. Occupational health nurses in Finland participate in continuing education at least every 5 years. In 1991 the option of specializing in Occupational Health as a part of the Masters of Health Care program became available at universities.

Occupational health nursing based on self care theory.

The study's purpose was that the nurses would understand self care theory and theoretical concepts in the background of nursing practice. At the beginning of the research the nurses set the clients' target and solved the clients' problems. As the research progressed the nurses let the clients set their targets. The nurses let the client make a decision. As the research progressed, the nurses began to understand the meaning of self care. The clients seemed to have been willing to take responsibility for their own self care.