Homocysteine and familial longevity: the Leiden Longevity Study

Homocysteine concentrations are a read-out of methionine metabolism and have been related to changes in lifespan in animal models. In humans, high homocysteine concentrations are an important predictor of age related disease. We aimed to explore the association of homocysteine with familial longevity by testing whether homocysteine is lower in individuals that are genetically enriched for longevity. We measured concentrations of total homocysteine in 1907 subjects from the Leiden Longevity Study consisting of 1309 offspring of nonagenarian siblings, who are enriched with familial factors promoting longevity, and 598 partners thereof as population controls. We found that homocysteine was related to age, creatinine, folate, vitamin B levels and medical history of hypertension and stroke in both groups (all p<0.001). However, levels of homocysteine did not differ between offspring enriched for longevity and their partners, and no differences in the age-related rise in homocysteine levels were found between groups (p for interaction 0.63). The results suggest that homocysteine metabolism is not likely to predict familial longevity.     

Familial longevity is marked by enhanced insulin sensitivity

Insulin resistance is a risk factor for various age-related diseases. In the Leiden Longevity study, we recruited long-lived siblings and their offspring. Previously, we showed that, compared to controls, the offspring of long-lived siblings had a better glucose tolerance. Here, we compared groups of offspring from long-lived siblings and controls for the relation between insulin and glucose in nonfasted serum (n = 1848 subjects) and for quantitation of insulin action using a two-step hyperinsulinemic-euglycemic clamp (n = 24 subjects). Groups of offspring and controls were similar with regard to sex distribution, age, and body mass index. We observed a positive bi-phasic linear relationship between ln (insulin) levels and nonfasted glucose with a steeper slope from 10.7mU L(-1) insulin onwards in controls compared to offspring (P = 0.02). During the clamp study, higher glucose infusion rate was required to maintain euglycemia during high-dose insulin infusion (P = 0.036) in offspring, reflecting higher whole-body insulin sensitivity. After adjustment for sex, age, and fat mass, the insulin-mediated glucose disposal rate (GDR) was higher in offspring than controls (42.5 ± 2.7 vs. 33.2 ± 2.7 micromol kg(-1) min(-1) , mean ± SE, P = 0.025). The insulin-mediated suppression of endogenous glucose production and lipolysis did not differ between groups (all P > 0.05). Furthermore, GDR was significantly correlated with the mean age of death of the parents. In conclusion, offspring from long-lived siblings are marked by enhanced peripheral glucose disposal. Future research will focus on identifying the underlying biomolecular mechanisms, with the aim to promote health in old age.     

Brain tissue volumes in familial longevity: the Leiden Longevity Study

Atrophy is one of the major age‐related changes in the brain. The absence of brain atrophy in elderly individuals reflects deceleration in the process of biological aging. Moreover, results from human twin studies suggest a large genetic influence on the variance of human brain tissue volumes. To investigate the association of brain volumes with exceptional longevity, we tested whether middle‐aged to elderly offspring of nonagenarian siblings have larger brain volumes than their spouses using magnetic resonance imaging. No differences in whole brain, gray matter and white matter volume were found. These brain volumes were associated with chronological age in offspring and control subjects (all P < 0.001). Left amygdalar volume of the offspring was larger (P = 0.03) compared with control subjects [mean volume offspring (cm³) (95% confidence interval, CI) = 1.39 (1.36–1.42), mean volume control subjects (cm³) (95% CI) = 1.32 (1.29–1.35)]. Association of left amygdalar volume with familial longevity was particularly pronounced when offspring with the oldest long‐lived parent were compared with control subjects (P = 0.01). Amygdalar volumes were not associated with chronological age in both groups. Our findings suggest that the observed association of a larger left amygdalar volume with familial longevity is not caused by a relative preservation of the left amygdala during the course of aging but most likely a result of early development caused by a genetic familial trait.     

Familial Longevity Is Marked by Better Cognitive Performance at Middle Age: The Leiden Longevity Study

Background

Decline in cognitive performance is a highly prevalent health condition in elderly. We studied whether offspring of nonagenarian siblings with a familial history of longevity, perform better on cognitive tests compared to their partners as controls. This is relevant since it could provide insights into determinants underlying decline in cognitive performance.

Methods

Cross-sectional analysis within the longitudinal cohort of the Leiden Longevity Study consisting of middle-aged offspring of nonagenarian siblings together with their partners (n = 500, mean age (SD) 66.3 (6.1) and 65.7 (7.2) years, respectively) as controls. Memory function, attention and processing speed were tested using the 15-Picture Learning Test, Stroop test and Digit Symbol Substitution Test. Data were analyzed with regression adjusted for age, gender, years of education and additionally for diabetes mellitus, cardiovascular diseases, alcohol use, smoking, inflammatory markers and apolipoprotein E genotype. Robust standard errors were used to account for familial relationships among the offspring.

Results

Cognitive performance was worse at higher calendar age (p<0.001, all except Stroop test part 1). The offspring performed better compared to their partners on trial 3 (p = 0.005), the immediate (p = 0.016) and delayed (p = 0.004) recall of the 15-Picture Learning Test as well as on the interference and combined interference score of the Stroop test (p = 0.014 and p = 0.036, respectively) in the fully adjusted model. The difference between offspring and partners was estimated to be more than three years according to the observed difference in calendar age.

Conclusions

Offspring of nonagenarian siblings with a familial history of longevity have better cognitive performance compared to the group of their partners of comparable age. This effect is independent of age-related diseases and known possible confounders. Possible explanations might be differences in subclinical vascular pathology between both groups.     

Levels of 25-hydroxyvitamin D in familial longevity: the Leiden Longevity Study

Background:

Low levels of 25(OH) vitamin D are associated with various age-related diseases and mortality, but causality has not been determined. We investigated vitamin D levels in the offspring of nonagenarians who had at least one nonagenarian sibling; these offspring have a lower prevalence of age-related diseases and a higher propensity to reach old age compared with their partners.

Methods:

We assessed anthropometric characteristics, 25(OH) vitamin D levels, parathyroid hormone levels, dietary vitamin D intake and single nucleotide polymorphisms (SNPs) associated with vitamin D levels. We included offspring (n = 1038) of nonagenarians who had at least one nonagenarian sibling, and the offsprings’ partners (n = 461; controls) from the Leiden Longevity Study. We included age, sex, body mass index, month during which blood sampling was performed, dietary and supplemental vitamin D intake, and creatinine levels as possible confounding factors.

Results:

The offspring had significantly lower levels of vitamin D (64.3 nmol/L) compared with controls (68.4 nmol/L; p = 0.002), independent of possible confounding factors. There was no difference in the levels of parathyroid hormone between groups. Compared with controls, the offspring had a lower frequency of a genetic variant in the CYP2R1 gene (rs2060793) (p = 0.04). The difference in vitamin D levels between offspring and controls persisted over the 2 most prevalent genotypes of this SNP.

Interpretation:

Compared with controls, the offspring of nonagenarians who had at least one nonagenarian sibling had a reduced frequency of a common variant in the CYP2R1 gene, which predisposes people to high vitamin D levels; they also had lower levels of vitamin D that persisted over the 2 most prevalent genotypes. These results cast doubt on the causal nature of previously reported associations between low levels of vitamin D and age-related diseases and mortality.Vitamin D plays a critical role in bone formation,¹ immune cell differentiation, and in the inhibition of proliferation and angiogenesis in cancer.² Low serum levels of 25-hydroxyvitamin D3 (25[OH] vitamin D) are associated with increased mortality, cardiovascular disease, diabetes mellitus, cancer, multiple sclerosis, allergy, asthma, infection, depression, mental illness and musculoskeletal pain.^3–6 However, because of design limitations, previous studies have not been able to infer causality.⁷ A meta-analysis that included 51 studies and more than 30 000 individuals in each treatment group showed no reduction in all-cause or cardiovascular mortality with vitamin D supplementation,⁸ suggesting that low serum levels of vitamin D are a consequence rather than a cause of disease.Vitamin D status is routinely assessed by measurement of serum concentration of the prohormone 25(OH) vitamin D, which is the most stable and abundant metabolite of vitamin D in circulation. In the current study, we investigated the association between serum levels of 25(OH) vitamin D and familial longevity. We assessed vitamin D levels in the offspring of nonagenarian siblings whom we have previously shown to have a lower prevalence of cardiovascular diseases and lower mortality compared to age- and environmental-matched controls.^9,10 We also assessed whether genetic variation in 3 genes¹¹ was associated with 25(OH) vitamin D levels among the offspring of nonagenarians who had at least 1 nonagenarian sibling, and their partners.     

Menstrual cycle variation in spatial ability: relation to salivary cortisol levels

This study examined whether menstrual cycle phase was associated with performance on the Primary Mental Abilities Test of Spatial Relations, a test of mental rotation, in undergraduate students (N = 82). As cortisol levels also vary across the menstrual cycle under conditions of stress and influence cognitive performance, saliva samples were obtained before and after the test session to examine whether cortisol levels were related to between- and within-group differences in spatial performance. Men scored higher on the spatial test than all the groups of women, although the difference between men and women in the menstrual phase was not significant. Women in the luteal phase scored lower than the menstrual, follicular, and oral contraceptive user groups of women. There were no sex or menstrual cycle differences in cortisol levels, and no association between cortisol levels and spatial performance. The poorer performance of women in the luteal phase was not related to differences in ratings of perceived stress, perceived success on the test, or mood. Although menstrual cycle phase accounted for a significant proportion of the variance (15%) in performance on the spatial test, this does not explain why men outperformed women regardless of the phase of the cycle. Thus, there are clearly several other variables, sociocultural and physiological, involved in mediating individual differences in spatial performance.     

Familial longevity is characterized by high circadian rhythmicity of serum cholesterol in healthy elderly individuals

The biological clock, whose function deteriorates with increasing age, determines bodily circadian (i.e. 24h) rhythms, including that of cholesterol metabolism. Dampening of circadian rhythms has been associated with aging and disease. Therefore, we hypothesized that individuals with a familial predisposition for longevity have a higher amplitude circadian serum cholesterol concentration rhythm. The aim of this study was to investigate circadian rhythmicity of serum cholesterol concentrations in offspring of nonagenarian siblings and their partners. Offspring from nonagenarian siblings (n = 19), and their partners as controls (n = 18), were recruited from the Leiden Longevity Study. Participants (mean age 65 years) were studied in a controlled in‐hospital setting over a 24‐h period, receiving three isocaloric meals at 9:00 h, 12:00 h and 18:00 h. Lights were off between 23:00 h and 8:00 h. Serum total cholesterol (TC), HDL cholesterol (HDL‐C), non‐HDL‐C and triglycerides (TG) were determined every 30 min over a 24‐h period. Serum TC concentrations were higher during day than during night in offspring (5.2 vs. 4.7 mm , P < 0.001) and in controls (5.3 vs. 5.0 mm , P < 0.001). The difference in TC concentrations between day and night tended to be greater in offspring than in controls (0.5 vs. 0.3 mm , P = 0.109), reaching statistical significance in females (P = 0.045). Notably, the day–night serum differences in non‐HDL‐C were twofold greater in offspring than in controls (0.43 vs. 0.21 mm , P = 0.044) and most explicit in females (0.53 vs. 0.22, P = 0.078). We conclude that familial longevity is characterized by a high circadian rhythmicity of non‐HDL‐C in healthy elderly offspring from nonagenarian siblings.     

Longevity of orders is related to the longevity of their constituent genera rather than genus richness

Longevity of a taxonomic group is an important issue in understanding the dynamics of evolution. In this respect a key observation is that genera, families or orders can each be assigned a characteristic average lifetime (Van Valen in Evol Theory 1:1–30, 1973). Using the fossil marine animal genera database (Sepkoski in Bull Am Paleontol 363, pp 563, 2002) we here examine the relationship between longevity of a higher taxonomic group (orders) and the longevity of its lower taxonomic groups (genera). We find insignificant correlation between the size of an order and its longevity, whereas we observe large correlation between the lifetime of an order and the lifetime of its constituent genera. These observations suggest that longevity of taxonomic groups is heritable intrinsically or on the grounds of environmental preferences.     

Human pancreatic polypeptide has a marked diurnal rhythm that is affected by ageing and is associated with the gastric TFF2 circadian rhythm

Normal circadian variations in vasoactive intestinal polypeptide, somatostatin, cholecystokinin and pancreatic polypeptide were measured to determine if these alter with aging and to identify gastrointestinal regulatory hormones that might control the dramatic diurnal variation in the gastric cytoprotective trefoil protein TFF2. Plasma pancreatic polypeptide concentrations showed a marked diurnal rhythm (p < 0.0001). Basal and postprandial pancreatic polypeptide concentrations increased with age (p < 0.01). The timing of the diurnal rhythm was consistent with pancreatic polypeptide inhibiting TFF2 secretion and there was a negative association between pancreatic polypeptide and TFF2 concentrations (p < 0.002). The much higher pancreatic polypeptide concentrations in older people will induce increased satiety that may contribute to 'anorexia of ageing'. These results identify potential therapies for treatment of gastric mucosal morbidity and age-associated loss of appetite.     

Effect of erythropoietin levels on mortality in old age: the Leiden 85-plus Study

Background

The production of erythropoietin is triggered by impaired oxygen delivery to the kidney, either because of anemia or hypoxemia. High erythropoietin levels have been shown to predict the risk of death among patients with chronic heart failure. We investigated the prognostic value of elevated erythropoietin levels on mortality among very elderly people in the general population.

Methods

The Leiden 85-plus Study is a population-based prospective follow-up study involving 599 people aged 85 years in Leiden, the Netherlands, enrolled between September 1997 and September 1999. Erythropoietin levels were determined at age 86. For this analysis, we included 428 participants with a creatinine clearance of at least 30 mL/min. Mortality data, recorded until Feb. 1, 2008, were obtained from the municipal registry.

Results

During follow-up, 324 (75.7%) participants died. Compared with participants whose erythropoietin levels were in the lowest tertile (reference group), those whose levels were in the middle tertile had a 25% increased risk of death (hazard ratio [HR] 1.25, 95% confidence interval [CI] 0.95–1.64), and those whose levels were in the highest tertile had a 73% increased risk (HR 1.73, 95% CI 1.32–2.26) (p value for trend < 0.01). The association between erythropoietin levels and mortality remained largely unchanged after we adjusted for sex, creatinine clearance, hemoglobin level, comorbidity, smoking status and C-reactive protein level, and was similar for deaths from cardiovascular and noncardiovascular causes.

Interpretation

Among people aged 85 years and older, elevated erythropoietin levels were associated with an increased risk of death, independent of hemoglobin levels.Decreased oxygen availability in the kidney triggers the peritubular capillary lining cells within the kidney to produce erythropoietin — the principal regulator of red blood cell mass.¹ Impaired oxygen delivery to the kidney can result from various pathophysiologic mechanisms, such as anemia, hypoperfusion due to renal arteriosclerosis, decreased renal blood flow or heart failure, and decreased oxygen saturation due to diseases such as chronic obstructive pulmonary disease.^1–5Studies involving patients with chronic heart failure have shown that high erythropoietin levels predict poor survival.^6–11 However, whether this prognostic value is limited to patients with heart failure or whether it is generalizable to very elderly people regardless of specific pathologic conditions has not been studied. We conducted this study to investigate whether high levels of erythropoietin predict mortality among very elderly people in the general population.     

Diurnal salivary cortisol is associated with body mass index and waist circumference: The multiethnic study of atherosclerosis

Objective:

Neuroendocrine abnormalities, such as activation of the hypothalamic‐pituitary‐adrenal (HPA) axis, are associated with obesity; however, few large‐scale population‐based studies have examined HPA axis and markers of obesity. We examined the cross‐sectional association of the cortisol awakening response (CAR) and diurnal salivary cortisol curve with obesity.

Design and Methods:

The Multiethnic Study of Atherosclerosis Stress Study includes 1,002 White, Hispanic, and Black men and women (mean age 65 ± 9.8 years) who collected up to 18 salivary cortisol samples over 3 days. Cortisol profiles were modeled using regression spline models that incorporated random parameters for subject‐specific effects. Cortisol curve measures included awakening cortisol, CAR (awakening to 30‐min postawakening), early decline (30 min to 2‐h postawakening), late decline (2‐h postawakening to bedtime), and the corresponding areas under the curve (AUC). Body mass index (BMI) and waist circumference (WC) were used to estimate adiposity.

Results:

For the entire cohort, both BMI and WC were negatively correlated with awakening cortisol (P < 0.05), AUC during awakening rise, and early decline and positively correlated to the early decline slope (P < 0.05) after adjustments for age, race/ethnicity, gender, diabetes status, socioeconomic status, β‐blockers, steroids, hormone replacement therapy, and smoking status. No heterogeneities of effects were observed by gender, age, and race/ethnicity.

Conclusions:

Higher BMI and WC are associated with neuroendocrine dysregulation, which is present in a large population sample, and only partially explained by other covariates.     

Familial dysautonomia is caused by mutations of the IKAP gene

The defective gene DYS, which is responsible for familial dysautonomia (FD) and has been mapped to a 0.5-cM region on chromosome 9q31, has eluded identification. We identified and characterized the RNAs encoded by this region of chromosome 9 in cell lines derived from individuals homozygous for the major FD haplotype, and we observed that the RNA encoding the IkappaB kinase complex-associated protein (IKAP) lacks exon 20 and, as a result of a frameshift, encodes a truncated protein. Sequence analysis reveals a T-->C transition in the donor splice site of intron 20. In individuals bearing a minor FD haplotype, a missense mutation in exon 19 disrupts a consensus serine/threonine kinase phosphorylation site. This mutation results in defective phosphorylation of IKAP. These mutations were observed to be present in a random sample of Ashkenazi Jewish individuals, at approximately the predicted carrier frequency of FD. These findings demonstrate that mutations in the gene encoding IKAP are responsible for FD.     

Influence of pregnancy on diurnal and seasonal changes in cortisol, T3 and T4 levels in the mare blood serum.

1. The diurnal changes in the level of total protein, cortisol, T3 and T4 were studied in four barren and four pregnant standard-bred mares, kept and examined under the same conditions. 2. Blood samples were taken every 4 hr. for one day each month, throughout one year. 3. In barren mares, a diurnal rhythm in cortisol level (acrophase at 0530 hr in summer and at 0830 hr in winter) and in T3 level (acrophase at 1330 hr in summer and at 1800 hr in winter) was found. 4. In pregnant mares, a diurnal rhythm in cortisol level only till 5th month of pregnancy was observed. 5. A diurnal rhythm in T3 level was found throughout the pregnancy, with acrophase always at 1400 hr. 6. No diurnal rhythm in the total protein content and in the T4 level was observed. 7. In both groups of mares the seasonal cyclicity in T3 and T4 levels were found. A seasonal cyclicity in cortisol level was found only in pregnant mares. 8. Pregnancy abolished seasonal cyclicity in total protein and showed it in cortisol level. 9. Pregnancy in mares modifies diurnal rhythms as well as seasonal cycles in secretion and metabolism of the hormones studied.     

Racial and ethnic differences in diurnal cortisol rhythms in preadolescents: the role of parental psychosocial risk and monitoring

Racial/ethnic minorities experience persistent health disparities due in part to their exposure to chronic SES and psychosocial risk. The hypothalamic-pituitary-adrenal axis and its hormonal end product, cortisol, are believed to mediate the associations between chronic stress and poor health. In this study, racial/ethnic differences in diurnal salivary cortisol rhythms in 179 preadolescent youths and the contributing roles of SES risk, psychosocial risk, perceived discrimination, harsh parenting, and parental monitoring were examined. The analyses revealed racial/ethnic differences in diurnal cortisol rhythms, with African Americans having significantly flatter morning-to-evening cortisol slopes than Caucasians and with Latinos having significantly lower evening cortisol levels than Caucasians. Greater psychosocial risk and less parental monitoring were associated with flatter cortisol slopes. Racial/ethnic differences on the cortisol measures persisted when controlling for SES, psychosocial risk, and parenting quality. The need to assess chronic risk across the lifespan and disentangle possible genetic from environmental contributors is discussed.     

A study of diurnal changes in cortisol and glucose levels and FDPA activity in foals during the first 13 weeks of life and in their lactating mothers

1. In six standard-bred mares and their foals diurnal changes in the cortisol and glucose levels and in FDPA activity were studies for 13 weeks of foal life. 2. In the cortisol level diurnal rhythm was found in the 3rd, 7th and 11th week of foal life and in the 11th week of lactation in mares. 3. In mares the mean diurnal cortisol level changed from 32 ng/ml in the first week to 57 in the 11th week and in foals from 24 in the first week to 16 ng/ml in the 11th week. 4. In the glucose level no diurnal rhythm was observed. 5. In mares the mean diurnal glucose level after parturition was about 58 mg/100 ml and increased to 83-85 mg/100 ml. In foals it changed from 105 to 128 mg/100 ml. 6. In the activity of FDPA no diurnal rhythm was observed. 7. The mean activity of FDPA changed in mares from 3.3 to 4.4 U and in foals from 5.7 to 7.5 U.     

Cytomegalovirus seropositivity is associated with glucose regulation in the oldest old. Results from the Leiden 85-plus Study

ABSTRACT: BACKGROUND: Cytomegalovirus (CMV) infection has been reported to contribute to the pathogenesis of type 1 diabetes and post-transplantation diabetes. However, CMV infection has not been evaluated as a possible risk factor for type 2 diabetes. Our aim was to investigate potential associations between CMV seropositivity, CMV IgG antibody level and glucose regulation in the oldest old. RESULTS: CMV seropositive subjects were more likely to have type 2 diabetes (17.2% vs 7.9%, p = 0.016), had a higher level of HbA1c (p = 0.014) and higher non-fasting glucose (p = 0.024) in the oldest olds. These associations remained significant after adjustment for possible confounders. CMV IgG antibody level was not significantly associated with glucose regulation (all p > 0.05). CONCLUSIONS: In the oldest old, CMV seropositivity is significantly associated with various indicators of glucose regulation. This finding suggests that CMV infection might be a risk factor for the development of type 2 diabetes in the elderly.     

Stress reactions in response to the procedure of LHRH tests as measured by salivary and serum cortisol and psychological variables

Serum and salivary cortisol release and mood ratings were measured in response to the procedure of LHRH tests in 12 medical students professionally familiarized with venipunctures (familiarized subgroup) and in 5 non-medical students (unfamiliarized subgroup). There were significant differences in cortisol and mood ratings between both groups. Only the unfamiliarized subgroup showed significant increases of cortisol and higher values for tension and anxiety. Relations between cortisol, tension and anxiety confirm an effect of experimental stress on the secretion of cortisol during the procedure of LHRH tests. Novelty and subjective mood states appear to have predictive values with regard to stress-induced endocrine reactions.     

Induction of cytoprotective pathways is central to the extension of lifespan conferred by multiple longevity pathways

Many genetic and physiological treatments that extend lifespan also confer resistance to a variety of stressors, suggesting that cytoprotective mechanisms underpin the regulation of longevity. It has not been established, however, whether the induction of cytoprotective pathways is essential for lifespan extension or merely correlated. Using a panel of GFP-fused stress response genes, we identified the suites of cytoprotective pathways upregulated by 160 gene inactivations known to increase Caenorhabditis elegans longevity, including the mitochondrial UPR (hsp-6, hsp-60), the ER UPR (hsp-4), ROS response (sod-3, gst-4), and xenobiotic detoxification (gst-4). We then screened for other gene inactivations that disrupt the induction of these responses by xenobiotic or genetic triggers, identifying 29 gene inactivations required for cytoprotective gene expression. If cytoprotective responses contribute directly to lifespan extension, inactivation of these genes would be expected to compromise the extension of lifespan conferred by decreased insulin/IGF-1 signaling, caloric restriction, or the inhibition of mitochondrial function. We find that inactivation of 25 of 29 cytoprotection-regulatory genes shortens the extension of longevity normally induced by decreased insulin/IGF-1 signaling, disruption of mitochondrial function, or caloric restriction, without disrupting normal longevity nearly as dramatically. These data demonstrate that induction of cytoprotective pathways is central to longevity extension and identify a large set of new genetic components of the pathways that detect cellular damage and couple that detection to downstream cytoprotective effectors.