Functional analysis of Aarf domain-containing kinase 1 in Drosophila melanogaster.

BACKGROUND: The ADCK proteins are predicted mitochondrial kinases. Most studies of these proteins have focused on the Abc1/Coq8 subfamily, which contributes to Coenzyme Q biosynthesis. In contrast, little is known about ADCK1 despite its evolutionary conservation in yeast, Drosophila, Caenorhabditis elegans and mammals. RESULTS: We show that Drosophila ADCK1 mutants die as second instar larvae with double mouth hooks and tracheal breaks. Tissue-specific genetic rescue and RNAi studies show that ADCK1 is necessary and sufficient in the trachea for larval viability. In addition, tracheal-rescued ADCK1 mutant adults have reduced lifespan, are developmentally delayed, have reduced body size, and normal levels of basic metabolites. CONCLUSION: The larval lethality and double mouth hooks seen in ADCK1 mutants are often associated with reduced levels of the steroid hormone ecdysone, suggesting that this gene could contribute to controlling ecdysone levels or bioavailability. Similarly, the tracheal defects in these animals could arise from defects in intracellular lipid trafficking. These studies of ADCK1 provide a new context to define the physiological functions of this poorly understood member of the ADCK family of predicted mitochondrial proteins.

Physiological responding to stress in middle-aged males enriched for longevity: a social stress study.

Individuals enriched for familial longevity display a lower prevalence of age-related diseases, such as cardiovascular- and metabolic diseases. Since these diseases are associated with stress and increased cortisol levels, one of the underlying mechanisms that may contribute to healthy longevity might be a more adaptive response to stress. To investigate this, male middle-aged offspring from long-lived families (n = 31) and male non-offspring (with no familial history of longevity) (n = 26) were randomly allocated to the Trier Social Stress Test or a control condition in an experimental design. Physiological (cortisol, blood pressure, heart rate) and subjective responses were measured during the entire procedure. The results showed that Offspring had lower overall cortisol levels compared to Non-offspring regardless of condition, and lower absolute cortisol output (AUCg) during stress compared to Non-Offspring, while the increase (AUCi) did not differ between groups. In addition, systolic blood pressure in Offspring was lower compared to Non-offspring during the entire procedure. At baseline, Offspring had significantly lower systolic blood pressure and reported less subjective stress than Non-offspring and showed a trend towards lower heart rate. Offspring from long-lived families might thus be less stressed prior to potentially stressful events and consequently show overall lower levels in physiological responses. Although attenuated physiological responding cannot be ruled out, lower starting points and a lower peak level in physiological responding when confronted with an actual stressor, might already limit damage due to stress over a lifetime. Lower physiological responding may also contribute to the lower prevalence of cardiovascular diseases and other stress-related diseases in healthy longevity.

A Chromosome 13 locus is associated with male-specific mortality in mice.

BACKGROUND AND AIM: Mortality is a highly complex trait influenced by a wide array of genetic factors. METHODS: We examined a population of 1200 mice that were F2 generation offspring of a 4-way reciprocal cross between C57BL6/J and DBA2/J strains. Animals were sacrificed at age 200, 500, or 800 days and genotyped at 96 markers. The 800 days old cohort, which were the survivors of a much larger breeding group, were examined for enriched frequency of alleles that benefit survival and depletion of alleles that reduce survival. RESULTS: Loci on Chr 13 in males and on Chr X in females were significantly distorted from Mendelian expectations, even after conservative correction for multiple testing. DBA2/J alleles between 35 and 80 Mb on Chr 13 were underrepresented in the age 800 male animals. D2 genotypes in this region were also associated with premature death during behavioral testing. Furthermore, confirmatory analysis showed BXD recombinant inbred strains carrying the D2 alleles in this region had shorter median survival. Exploration of available pathology data indicated that a syndrome involving dental malocclusions, pancreatic islet hypertrophy, and kidney lipidosis may have mediated the effects of DBA alleles on mortality specifically in male mice. The heterozygote advantage locus on the X Chr was not found to be associated with any pathology. CONCLUSIONS: These results suggest a novel locus influencing survival in the B6/D2 genetic background, perhaps via a metabolic disorder that emerges by 200 days of age in male animals.

New clinical findings on the longevity gene in disease, health, & longevity: Sirtuin 1 often decreases with advanced age & serious diseases in most parts of the human body, while relatively high & constant Sirtuin 1 regardless of age was first found in the hippocampus of supercentenarians.

The expression of the longevity gene, Sirtuin 1, was non-invasively measured using Electro-Magnetic Field (EMF) resonance phenomenon between a known amount of polyclonal antibody of the C-terminal of Sirtuin 1 & Sirtuin 1 molecule inside of the body. Our measurement of over 100 human adult males and females, ranging between 20-122 years old, indicated that the majority of subjects had Sirtuin 1 levels of 5-10 pg BDORT units in most parts of the body. When Sirtuin 1 was less than 1 pg, the majority of the people had various degrees of tumors or other serious diseases. When Sirtuin 1 levels were less than 0.25 pg BDORT units, a high incidence of AIDS was also detected. Very few people had Sirtuin 1 levels of over 25 pg BDORT units in most parts of the body. We selected 7 internationally recognized supercentenarians who lived between 110-122 years old. To our surprise, most of their body Sirtuin 1 levels were between 2.5-10 pg BDORT units. However, by evaluating different parts of the brain, we found that both sides of the Hippocampus had a much higher amount of Sirtuin 1, between 25-100 pg BDORT units. With most subjects, Sirtuin 1 was found to be higher in the Hippocampus than in the rest of the body and remains relatively constant regardless of age. We found that Aspartame, plastic eye contact lenses, and asbestos in dental apparatuses, which reduce normal cell telomeres, also significantly reduce Sirtuin 1. In addition, we found that increasing normal cell telomere by electrical or mechanical stimulation of True ST-36 increases the expression of the Sirtuin 1 gene in people in which expression is low. This measurement of Sirtuin 1 in the Hippocampus has become a reliable indicator for detecting potential longevity of an individual.

Metformin, valproic acid, and starvation induce seizures in a patient with partial SLC13A5 deficiency: a case of pharmaco-synergistic heterozygosity.

SLC13A5/NaCT is a sodium-coupled citrate transporter expressed in the plasma membrane of the liver, testis, and brain. In these tissues, SLC13A5 has important functions in the synthesis of fatty acids, cholesterol, and neurotransmitters. In recent years, patients homozygous for recessive mutations in SLC13A5, known as SLC13A5 deficiency [early infantile epileptic encephalopathy-25 (EIEE-25)], exhibit severe global developmental delay, early-onset intractable seizures, spasticity, and amelogenesis imperfecta affecting tooth development. Although the pathogenesis of SLC13A5 deficiency remains not clearly understood, cytoplasmic citrate deficits, decreased energy status in neurons, and citrate-zinc chelation are hypothesized to explain the neurological deficits. However, no study has examined the possibility of specific pharmacological drugs and/or lifestyle changes synergizing with heterozygosity of SLC13A5 deficiency to increase the risk of EIEE-25 clinical phenotype. Here, we report on a heterozygous SLC13A5-deficient patient who demonstrated evidence of pharmaco-synergistic heterozygosity upon administration of metformin, valproic acid, and starvation. The report illustrates the importance of careful consideration of the potential adverse effects of specific pharmacological treatments in patients with heterozygosity for disease-causing recessive mutations in SLC13A5.

Getting long in the tooth: a strong positive correlation between canine size and heterozygosity in Antarctic fur seals Arctocephalus gazella.

Most studies of heterozygosity-fitness correlations (HFCs) in natural populations relate to fitness traits expressed early in life, whereas traits that are often more difficult to measure such as longevity and adult body size remain elusive. Teeth provide a window on an individual's life history, allowing the reliable estimation of both age and body size. Consequently, we collected paired upper canine teeth and tissue samples from 84 adult male Antarctic fur seals Arctocephalus gazella that died of natural causes at Bird Island, South Georgia. Tooth size is a good predictor of skull and body size both within and across taxa, and we similarly find a strong relationship with skull size in our species. In turn, tooth size is itself predicted strongly by genetic heterozygosity estimated using 9 microsatellites. With only 9 loci, the exact mechanisms involved remain unclear, although the observed pattern appears largely attributable to a small subset of loci, suggesting that associative overdominance rather than inbreeding depression provides the proximate mechanism. In addition, locating these markers in the dog genome reveals proximity to genes involved with fat metabolism and growth. Our study illustrates how canine teeth, and potentially other structures such as tympano-periotic bone, waxy inner earplugs, or otoliths, may be used to explore links between genetic variation and important life-history traits in free-ranging vertebrate populations.

Evaluation of F1 cows sired by Brahman, Boran, and Tuli bulls for reproductive, maternal, and cow longevity traits.

This study evaluated reproductive, maternal performance, and longevity traits of 143 F1 cows sired by Brahman (Br), Boran (Bo), or Tuli (T) bulls from Angus or Hereford cows from 1994 to 2011. Cow traits were measured at 7 yr of age in 1999 and 2000 for 1992- and 1993-born cows, respectively. From 2004 to 2010, excluding 2008, incisor condition (solid, broken, smooth) scores were assigned to cows remaining in production; scores were evaluated with two models. Broken and solid mouths were each assigned a score of "1" and smooth assigned "0"; Br-sired (0.76) and Bo-sired cows (0.71) had higher scores (P < 0.05) than T-sired cows (0.54). When solid mouths were scored 1 and smooth and broken scored 0, Br-sired cows (0.34) were higher than T-sired (0.01) (P < 0.05), and Bo-sired (0.23) cows were not different from either (P > 0.05). Age level of the cow within birth year was important for all modeled calf traits (P < 0.05). Birth weights were not different among cow inheritance (P > 0.05). Cow type influenced (P < 0.05) 205-d adjusted weaning weight of calves; Br-sired dams (228.1 ± 2.37 kg) produced the greatest weaning weight, followed by Bo-sired (213.7 ± 3.10 kg), and T-sired (201.6 ± 2.69 kg) dams (P < 0.05). Adjusted means for calving rate for Bo-sired (0.92 ± 0.02) cows were higher (P < 0.05) than Br-sired (0.86 ± 0.02) and T-sired (0.86 ± 0.02) cows. Adjusted mean weaning rate was greater (P < 0.05) for Bo-sired cows (0.86 ± 0.02) than for cows sired by Br (0.77 ± 0.02) bulls, but weaning rate for T-sired cows (0.80 ± 0.02) were similar (P > 0.05). Cow weight was greater (P < 0.05) for Br-sired cows (590.5 ± 8.35 kg) than for Bo-sired (505.8 ± 10.46 kg) or T-sired cows (508.5 ± 9.37 kg). BCS at weaning for 7-yr-old cows was similar (P = 0.08) for Br-sired and Bo-sired cows and lower for T-sired cows (P = 0.0005, condition scores 6.0, 6.3, and 5.8, respectively). Boran-sired cows were older when they were removed from the herd, on average (12.7 ± 0.74 y, P = 0.03) than T-sired (10.6 ± 0.61 y); Br-sired cow persistence was intermediate and not different (11.05 ± 0.60 y, P > 0.06) from the others. Boran-sired cows had higher calving and weaning rates and better mouth scores than the other groups; consequently, they had greater longevity as well. Boran-sired and T-sired cows were moderate in size and weighed less than Br-sired cows throughout the study. Tuli-sired cows weaned the lightest calves and had the most tooth deterioration as they aged.

Knockdown of the Drosophila FIG4 induces deficient locomotive behavior, shortening of motor neuron, axonal targeting aberration, reduction of life span and defects in eye development.

Mutations in Factor-Induced-Gene 4 (FIG4) gene have been identified in Charcot-Marie-Tooth disease type 4J (CMT4J), Yunis-Varon syndrome and epilepsy with polymicrogyria. FIG4 protein regulates a cellular abundance of phosphatidylinositol 3,5-bisphosphate (PI(3,5)P2), a signaling lipid on the cytosolic surface of membranes of the late endosomal compartment. PI(3,5)P2 is required for retrograde membrane trafficking from lysosomal and late endosomal compartments to the Golgi. However, it is still unknown how the neurodegeneration that occurs in these diseases is related to the loss of FIG4 function. Drosophila has CG17840 (dFIG4) as a human FIG4 homolog. Here we specifically knocked down dFIG4 in various tissues, and investigated their phenotypes. Neuron-specific knockdown of dFIG4 resulted in axonal targeting aberrations of photoreceptor neurons, shortened presynaptic terminals of motor neurons in 3rd instar larvae and reduced climbing ability in adulthood and life span. Fat body-specific knockdown of dFIG4 resulted in enlarged lysosomes in cells that were detected by staining with LysoTracker. In addition, eye imaginal disk-specific knockdown of dFIG4 disrupted differentiation of pupal ommatidial cell types, such as cone cells and pigment cells, suggesting an additional role of dFIG4 during eye development.

Inflammation, But Not Telomere Length, Predicts Successful Ageing at Extreme Old Age: A Longitudinal Study of Semi-supercentenarians.

To determine the most important drivers of successful ageing at extreme old age, we combined community-based prospective cohorts: Tokyo Oldest Old Survey on Total Health (TOOTH), Tokyo Centenarians Study (TCS) and Japanese Semi-Supercentenarians Study (JSS) comprising 1554 individuals including 684 centenarians and (semi-)supercentenarians, 167 pairs of centenarian offspring and spouses, and 536 community-living very old (85 to 99 years). We combined z scores from multiple biomarkers to describe haematopoiesis, inflammation, lipid and glucose metabolism, liver function, renal function, and cellular senescence domains. In Cox proportional hazard models, inflammation predicted all-cause mortality with hazard ratios (95% CI) 1.89 (1.21 to 2.95) and 1.36 (1.05 to 1.78) in the very old and (semi-)supercentenarians, respectively. In linear forward stepwise models, inflammation predicted capability (10.8% variance explained) and cognition (8(.)6% variance explained) in (semi-)supercentenarians better than chronologic age or gender. The inflammation score was also lower in centenarian offspring compared to age-matched controls with Δ (95% CI) = - 0.795 (- 1.436 to - 0.154). Centenarians and their offspring were able to maintain long telomeres, but telomere length was not a predictor of successful ageing in centenarians and semi-supercentenarians. We conclude that inflammation is an important malleable driver of ageing up to extreme old age in humans.

Heterosis for survival and dentition in Hereford, Angus, Shorthorn, and crossbred cows.

Records of 328 Angus, Hereford, Shorthorn, and first-cross cows produced from 1960 to 1963 were used to estimate heterosis (h) and individual (gI) and maternal (gM) average breed effects on cumulative survival (CS) by ages to 12 yr, longevity (L), and size (SI) and condition of incisors at 10 to 15 yr of age. Reasons for cow disposal were also studied. The 155 cows born in 1960 and 1961 first calved at 3 yr of age, and the 173 born in 1962 and 1963 first calved at 2 yr of age. Analyses for CS and L were done for both actual culling of cows open in two consecutive years (AC) and imposed culling of any open cow (IC). The model for CS and L included cow birth year-breed of cow's sire (Y-S), sires within Y-S, breed of dam (D), and D x Y-S. For SI, the model included breed of cow, year of recording, and quadratic effect of age. Linear contrasts were used to estimate h, gI, and gM for quantitative variables, and chi-square was used for discrete variables. Under both AC (P less than .05) and IC, crossbreds exceeded straightbreds in CS to 12 yr (16.7 and 8.5%) and L (1.36 and .99 yr). Estimates of gI and gM for CS and L were generally small, except for the low (P less than .05) gM of Shorthorn cows under IC. Mortality and culling for emaciation, cancer eye, or prolapse was less for crossbreds than for straightbreds and increased with age. Crossbreds had longer (P less than .01) and better (P less than .001) teeth than straightbreds. The longer productive life of crossbred cows reduces rearing costs for replacements and increases sales of calves and cull cows.

WormFarm: a quantitative control and measurement device toward automated Caenorhabditis elegans aging analysis.

Caenorhabditis elegans is a leading model organism for studying the basic mechanisms of aging. Progress has been limited, however, by the lack of an automated system for quantitative analysis of longevity and mean lifespan. To address this barrier, we developed 'WormFarm', an integrated microfluidic device for culturing nematodes. Cohorts of 30-50 animals are maintained throughout their lifespan in each of eight separate chambers on a single WormFarm polydimethylsiloxane chip. Design features allow for automated removal of progeny and efficient control of environmental conditions. In addition, we have developed computational algorithms for automated analysis of video footage to quantitate survival and other phenotypes, such as body size and motility. As proof-of-principle, we show here that WormFarm successfully recapitulates survival data obtained from a standard plate-based assay for both RNAi-mediated and dietary-induced changes in lifespan. Further, using a fluorescent reporter in conjunction with WormFarm, we report an age-associated decrease in fluorescent intensity of GFP in transgenic worms expressing GFP tagged with a mitochondrial import signal under the control of the myo-3 promoter. This marker may therefore serve as a useful biomarker of biological age and aging rate.

The western painted turtle genome, a model for the evolution of extreme physiological adaptations in a slowly evolving lineage.

BACKGROUND: We describe the genome of the western painted turtle, Chrysemys picta bellii, one of the most widespread, abundant, and well-studied turtles. We place the genome into a comparative evolutionary context, and focus on genomic features associated with tooth loss, immune function, longevity, sex differentiation and determination, and the species' physiological capacities to withstand extreme anoxia and tissue freezing. RESULTS: Our phylogenetic analyses confirm that turtles are the sister group to living archosaurs, and demonstrate an extraordinarily slow rate of sequence evolution in the painted turtle. The ability of the painted turtle to withstand complete anoxia and partial freezing appears to be associated with common vertebrate gene networks, and we identify candidate genes for future functional analyses. Tooth loss shares a common pattern of pseudogenization and degradation of tooth-specific genes with birds, although the rate of accumulation of mutations is much slower in the painted turtle. Genes associated with sex differentiation generally reflect phylogeny rather than convergence in sex determination functionality. Among gene families that demonstrate exceptional expansions or show signatures of strong natural selection, immune function and musculoskeletal patterning genes are consistently over-represented. CONCLUSIONS: Our comparative genomic analyses indicate that common vertebrate regulatory networks, some of which have analogs in human diseases, are often involved in the western painted turtle's extraordinary physiological capacities. As these regulatory pathways are analyzed at the functional level, the painted turtle may offer important insights into the management of a number of human health disorders.

Age relationships of postmortem observations in Portuguese Water Dogs.

A dog model has been used to evaluate histological changes arising from senescence. Autopsies of 145 Portuguese Water Dogs have been used to evaluate the individual and group "state of health" at time of death. For each dog, weights or dimensions of organs or tissues were obtained, together with histological evaluation of tissues. Twenty-three morphological metrics correlated significantly to age at death. Many of these involved muscles; others were associated with derivatives of embryonic foregut. The latter included lengths of the small intestine and trachea as well as weights of the stomach and some lung lobes. Nearly all of the dogs examined had histological changes in multiple tissues, ranging from two to 12 per dog. Associations among pathologies included inflammatory bowel disease with osteoporosis and dental calculus/periodontitis with atherosclerosis and amyloidosis. In addition, two clusters of histological changes were correlated to aging: hyperplasia, frequency of adenomas, and hemosiderosis constituted one group; inflammation, plasmacytic and lymphocytic infiltration, fibrosis, and atrophy, another. Heritability analysis indicated that many of the changes in tissue/organ morphology or histology could be heritable and possibly associated with IGF1, but more autopsies will be required to substantiate these genetic relationships.

Genetic relationship between longevity and objectively or subjectively assessed performance traits in sheep using linear censored models.

Genetic parameters of longevity in crossbred Mule ewes, and genetic relationships among longevity, growth, body composition, and subjectively assessed traits on Mule lambs and ewes have been estimated using Bayesian linear censored models. Additionally, the genetic associations between longevity and culling reasons were examined. Data comprised 1,797 observations of Mule ewes for longevity, culling reasons, growth, body composition, mouth scores, and type traits. Longevity was defined as the time (in years) from 2 yr of age (the age at first lambing of most ewes) to culling or death. Censored data (i.e., observations for which only the lower bound of the true longevity is known, such as when the animals are still alive) comprised 24% of all observations for longevity. Bivariate analyses were used to analyze the longevity of the ewe with each performance trait by fitting linear Bayesian models considering censored observations. Longevity was split into 3 different sub-traits: age at culling due to teeth/mouth conditions, age at culling due to udder conditions, and age at culling due to other culling reasons. These sub-traits and their aggregation into the overall trait of longevity were analyzed in a multiple-trait model. The heritability of longevity was moderate at 0.27, whereas heritabilities of the growth and body composition traits ranged from 0.11 for average of shoulder, loin, and gigot conformation to 0.36 for ewe BW at first premating. Mouth scores and type traits had heritabilities ranging from 0.13 for jaw position to 0.39 for fleece quality. All analyzed traits showed low genetic correlations with longevity, ranging from -0.20 for average conformation scores in live animals to 0.18 for tooth angle. Teeth/mouth conditions resulted in the greatest heritability (0.15) among the sub-traits based on the separate culling reasons. Genetic correlations between separate culling reasons were low to high (0.12 to 0.63 for teeth/mouth conditions with udder conditions and other culling reasons, respectively). Longevity may be preferred as a selection criterion because of (i) its moderate heritability compared with its component sub-traits based on specific culling reasons, and (ii) its moderate to high genetic correlation with these component sub-traits. The moderate heritability for longevity reflects the potential of this trait for genetic improvement, especially when longevity is based on clearly defined culling reasons.