A novel estimate of biological aging by multiple fitness tests is associated with risk scores for age-related diseases.

Introduction: Recent research highlights the need for a correct instrument for monitoring the individual health status, especially in the elderly. Different definitions of biological aging have been proposed, with a consistent positive association of physical activity and physical fitness with decelerated aging trajectories. The six-minute walking test is considered the current gold standard for estimating the individual fitness status in the elderly. Methods: In this study, we investigated the possibility of overcoming the main limitations of assessing fitness status based on a single measure. As a result, we developed a novel measure of fitness status based on multiple fitness tests. In 176 Sardinian individuals aged 51-80 years we collected the results of eight fitness tests to measure participants' functional mobility, gait, aerobic condition, endurance, upper and lower limb strength, and static and dynamic balance. In addition, the participants' state of health was estimated through validated risk scores for cardiovascular diseases, diabetes, mortality, and a comorbidity index. Results: Six measures contributing to fitness age were extracted, with TUG showing the largest contribution (beta = 2.23 SDs), followed by handgrip strength (beta = -1.98 SDs) and 6MWT distance (beta = -1.11 SDs). Based on fitness age estimates, we developed a biological aging measure using an elastic net model regression as a linear combination of the results of the fitness tests described above. Our newly developed biomarker was significantly associated with risk scores for cardiovascular events (ACC-AHA: r = 0.61; p = 0.0006; MESA: r = 0.21; p = 0.002) and mortality (Levine mortality score: r = 0.90; p = 0.0002) and outperformed the previous definition of fitness status based on the six-minute walking test in predicting an individual health status. Discussion: Our results indicate that a composite measure of biological age based on multiple fitness tests may be helpful for screening and monitoring strategies in clinical practice. However, additional studies are needed to test standardisation and to calibrate and validate the present results.

An epigenome-wide association study meta-analysis of educational attainment.

The epigenome is associated with biological factors, such as disease status, and environmental factors, such as smoking, alcohol consumption and body mass index. Although there is a widespread perception that environmental influences on the epigenome are pervasive and profound, there has been little evidence to date in humans with respect to environmental factors that are biologically distal. Here we provide evidence on the associations between epigenetic modifications-in our case, CpG methylation-and educational attainment (EA), a biologically distal environmental factor that is arguably among the most important life-shaping experiences for individuals. Specifically, we report the results of an epigenome-wide association study meta-analysis of EA based on data from 27 cohort studies with a total of 10 767 individuals. We find nine CpG probes significantly associated with EA. However, robustness analyses show that all nine probes have previously been found to be associated with smoking. Only two associations remain when we perform a sensitivity analysis in the subset of never-smokers, and these two probes are known to be strongly associated with maternal smoking during pregnancy, and thus their association with EA could be due to correlation between EA and maternal smoking. Moreover, the effect sizes of the associations with EA are far smaller than the known associations with the biologically proximal environmental factors alcohol consumption, body mass index, smoking and maternal smoking during pregnancy. Follow-up analyses that combine the effects of many probes also point to small methylation associations with EA that are highly correlated with the combined effects of smoking. If our findings regarding EA can be generalized to other biologically distal environmental factors, then they cast doubt on the hypothesis that such factors have large effects on the epigenome.

Early diagnosis of bladder cancer through the detection of urinary tyrosine-phosphorylated proteins.

BACKGROUND: A noninvasive, highly sensitive and specific urine test is needed for bladder cancer (BC) diagnosis and surveillance in addition to the invasive cystoscopy. We previously described the diagnostic effectiveness of urinary tyrosine-phosphorylated proteins (UPY) and a new assay (UPY-A) for their measurement in a pilot study. The aim of this work was to evaluate the performances of the UPY-A using an independent cohort of 262 subjects. METHODS: Urinary tyrosine-phosphorylated proteins were measured by UPY-A test. The area under ROC curve, cutoff, sensitivity, specificity and predictive values of UPY-A were determined. The association of UPY levels with tumour staging, grading, recurrence and progression risk was analysed by Kruskal-Wallis and Wilcoxon's test. To test the probability to be a case if positive at the UPY-A, a logistic test adjusted for possible confounding factor was used. RESULTS: Results showed a significant difference of UPY levels between patients with BC vs healthy controls. For the best cutoff value, 261.26 Standard Units (SU), the sensitivity of the assay was 80.43% and the specificity was 78.82%. A statistically significant difference was found in the levels of UPY at different BC stages and grades between Ta and T1 and with different risk of recurrence and progression. A statistically significant increased risk for BC at UPY-A ⩾261.26 SU was observed. CONCLUSIONS: The present study supplies important information on the diagnostic characteristics of UPY-A revealing remarkable performances for early stages and allowing its potential use for different applications encompassing the screening of high-risk subjects, primary diagnosis and posttreatment surveillance.

B-vitamins intake, DNA-methylation of One Carbon Metabolism and homocysteine pathway genes and myocardial infarction risk: the EPICOR study.

BACKGROUND AND AIMS: Several epidemiological studies highlighted the association between folate and B-vitamins low intake and cardiovascular diseases (CVD) risk. Contrasting results were reported on the relationship between folate intake and DNA-methylation. Folate and B-vitamins may modulate DNA-methylation of specific enzymes which are included in the One-Carbon Metabolism (OCM) and in the homocysteine (Hcy) pathways. The aim of the study was to evaluate whether DNA-methylation profiles of OCM and Hcy genes could modulate the myocardial infarction (MI) risk conferred by a low B-vitamins intake. METHODS AND RESULTS: Study sample (206 MI cases and 206 matched controls) is a case-control study nested in the prospective EPIC cohort. Methylation levels of 33 candidate genes where extracted by the whole epigenome analysis (Illumina-HumanMethylation450K-BeadChip). We identified three differentially methylated regions in males (TCN2 promoter, CBS 5'UTR, AMT gene-body) and two in females (PON1 gene-body, CBS 5'UTR), each of them characterized by an increased methylation in cases. Functional in silico analysis suggested a decreased expression in cases. A Recursively Partitioned Mixture Model cluster algorithm identified distinct methylation profiles associated to different MI risk: high-risk vs. low-risk methylation profile groups, OR = 3.49, p = 1.87 × 10(-)(4) and OR = 3.94, p = 0.0317 in males and females respectively (multivariate logistic regression adjusted for classical CVD risk factors). Moreover, a general inverse relationship between B-vitamins intake and DNA-methylation of the candidate genes was observed. CONCLUSIONS: Our findings support the hypothesis that DNA-methylation patterns in specific regions of OCM and Hcy pathways genes may modulate the CVD risk conferred by folate and B-vitamins low intake.

Lessons learned from an osteoporosis clinical trial in frail long-term care residents.

BACKGROUND: Although osteoporosis affects women of all ages, the impact is most pronounced in frail residents in long-term care. Nevertheless, few interventional trials have been performed in this population, and few data on therapeutic alternatives are available in this cohort. PURPOSE: We describe the challenges and lessons learned in developing and carrying out a trial in frail long-term-care residents. METHODS: The Zoledronic acid in frail Elders to STrengthen bone (ZEST) study was designed to examine the safety and efficacy of a single-dose therapy for osteoporosis in frail residents in long-term care in the Pittsburgh area. Women with osteoporosis who were 65 years of age and older and currently not on therapy were randomized in a blinded fashion to intravenous zoledronic acid or placebo. Follow-up of each participant was planned for 2 years. All participants received appropriate calcium and vitamin D supplementation. RESULTS: Seven hundred and thirty-three contacts were made with long-term care residents of nine participating facilities. Of 252 women screened, 181 were eligible, enrolled, and randomized. Multiple barriers to research in long-term-care facilities were encountered but overcome with direct communication, information sessions, in-service trainings, and social events. Lessons learned included designing the study in a manner that avoided placing an additional burden on an already overcommitted facility staff, a two-stage consent process to separate screening from randomization, and a flexible examination schedule to accommodate residents while obtaining the necessary outcome measurements. Furthermore, a mobile unit accessible to participants containing state-of-the-art dual x-ray absorptiometry (DXA), assessment for vertebral fractures, and phlebotomy equipment allows all assessments to be performed on-site at each facility. Serious adverse events are collected from affiliated hospitals in real time with a novel electronic surveillance system. LIMITATIONS: The major limitation is selection of outcomes that can be assessed at participating facilities and do not require transport of participants to hospitals or clinics. CONCLUSIONS: Clinical research for osteoporosis can be successfully and safely performed with frail residents in long-term care facilities. Lessons learned from this study may inform future investigations among frail elderly residents of these facilities.

Control of octopus arm extension by a peripheral motor program.

For goal-directed arm movements, the nervous system generates a sequence of motor commands that bring the arm toward the target. Control of the octopus arm is especially complex because the arm can be moved in any direction, with a virtually infinite number of degrees of freedom. Here we show that arm extensions can be evoked mechanically or electrically in arms whose connection with the brain has been severed. These extensions show kinematic features that are almost identical to normal behavior, suggesting that the basic motor program for voluntary movement is embedded within the neural circuitry of the arm itself. Such peripheral motor programs represent considerable simplification in the motor control of this highly redundant appendage.

Prey-handling behaviour of Octopus vulgaris (Mollusca, Cephalopoda) on Bivalve preys.

This study examines the ability of Octopus vulgaris to prey on bivalve molluscs showing that octopuses exhibited the same behavioural pattern irrespective of the prey species. When the initial pulling method did not result efficient octopuses skipped to drilling accompanied by an increase in the handling time. A complete behavioural repertoire of O. vulgaris in handling bivalve preys is also given.

Scopolamine impairs memory recall in Octopus vulgaris.

The involvement of the central cholinergic system in predatory performance, and on the recall of individual and observational memory in Octopus vulgaris was studied by treating the animals with the muscarinic antagonist scopolamine (2 mg/kg). The absence of the effects of the injection of scopolamine on blood circulation was also checked. Scopolamine did not affect the ability of octopuses to prey on live crabs. However, it interfered significantly with memory recall. In fact, the ability to solve the jar problem was impaired within the first hour after injection (short-term effects) and was only partially recovered after 24 h (long-term). Moreover, both individual and observational learning of a visual discrimination were significantly reduced at the short- and long-term testing. These results support a role of the cholinergic system in the processes of memory recall of O. vulgaris.

Patterns of arm muscle activation involved in octopus reaching movements.

The extreme flexibility of the octopus arm allows it to perform many different movements, yet octopuses reach toward a target in a stereotyped manner using a basic invariant motor structure: a bend traveling from the base of the arm toward the tip (Gutfreund et al., 1996a). To study the neuronal control of these movements, arm muscle activation [electromyogram (EMG)] was measured together with the kinematics of reaching movements. The traveling bend is associated with a propagating wave of muscle activation, with maximal muscle activation slightly preceding the traveling bend. Tonic activation was occasionally maintained afterward. Correlation of the EMG signals with the kinematic variables (velocities and accelerations) reveals that a significant part of the kinematic variability can be explained by the level of muscle activation. Furthermore, the EMG level measured during the initial stages of movement predicts the peak velocity attained toward the end of the reaching movement. These results suggest that feed-forward motor commands play an important role in the control of movement velocity and that simple adjustment of the excitation levels at the initial stages of the movement can set the velocity profile of the whole movement. A simple model of octopus arm extension is proposed in which the driving force is set initially and is then decreased in proportion to arm diameter at the bend. The model qualitatively reproduces the typical velocity profiles of octopus reaching movements, suggesting a simple control mechanism for bend propagation in the octopus arm.

The role of stimulus preexposure in problem solving by Octopus vulgaris.

Octopus vulgaris is able to open transparent glass jars closed with plastic plugs and containing live crabs. The decrease in performance times for removing the plug and seizing the prey with increasing experience of the task has been taken to indicate learning. However, octopuses' attack behaviors are typically slow and variable in novel environmental situations. In this study the role of preexposure to selected features of the problem-solving context was investigated. Although octopuses failed to benefit from greater familiarity with the training context or with selected elements of the task of solving the jar problem, the methodological strategies used are instructive in potentially clarifying the role of complex problem-solving behaviors in this species including stimulus preexposure and social learning.

Organization of octopus arm movements: a model system for studying the control of flexible arms.

Octopus arm movements provide an extreme example of controlled movements of a flexible arm with virtually unlimited degrees of freedom. This study aims to identify general principles in the organization of these movements. Video records of the movements of Octopus vulgaris performing the task of reaching toward a target were studied. The octopus extends its arm toward the target by a wave-like propagation of a bend that travels from the base of the arm toward the tip. Similar bend propagation is seen in other octopus arm movements, such as locomotion and searching. The kinematics (position and velocity) of the midpoint of the bend in three-dimensional space were extracted using the direct linear transformation algorithm. This showed that the bend tends to move within a single linear plane in a simple, slightly curved path connecting the center of the animal's body with the target location. Approximately 70% of the reaching movements demonstrated a stereotyped tangential velocity profile. An invariant profile was observed when movements were normalized for velocity and distance. Two arms, extended together in the same behavioral context, demonstrated identical velocity profiles. The stereotyped features of the movements were also observed in spontaneous arm extensions (not toward an external target). The simple and stereotypic appearance of the bend trajectory suggests that the position of the bend in space and time is the controlled variable. We propose that this strategy reduces the immense redundancy of the octopus arm movements and hence simplifies motor control.

Octopus vulgaris (Mollusca, Cephalopoda) as a model in behavioral pharmacology: a test of handling effects.

Despite the great interest of Cephalopods in learning studies, behavioral pharmacological experiments using these animals are scanty. The purpose of this study was to find an appropriate method of injection of substances for studying their effects on the behavior of octopuses. We injected into the branchial heart a known volume of seawater to test the effect of cold anaesthesia and of different types of manipulation on the predatory performance of Octopus vulgaris. An injection procedure that is simple, reliable, and does not require anaesthesia is proposed. The article also addresses ethical and manipulation requirements of modern behavioral pharmacology.

Lesions of the vertical lobe impair visual discrimination learning by observation in Octopus vulgaris.

We investigated whether lesions of the vertical lobe of the supraesophageal nervous mass of Octopus vulgaris impair discrimination learning acquired by observation of conspecific behavior. When tested alone, observer octopuses with about 50% of the vertical lobe removed showed a deficit in their learning by observation performance. The level of learning improved 24 h after the observational phase, when observer Octopus vulgaris showed a visual discriminatory performance significantly in agreement with the observed one. Control animals that had no brain tissue removed, did not show any impairment in the discriminative performance they had acquired vicariously. Removal of the vertical lobe of the octopus 'brain' has been reported to induce learning and memory deficits of visual discrimination in direct learning by conditioning experiments. Our findings support the conclusion that the removal of such brain center impairs short-term recall, but does not impair acquisition nor retention of 'observational' long-term memory.

Observational Learning in Octopus vulgaris.

Untrained Octopus vulgaris (observers) were allowed to watch conditioned Octopus(demonstrators) perform the task of selecting one of two objects that were presented simultaneously and differed only in color. After being placed in isolation, the observers, in a similar test, consistently selected the same object as did the demonstrators. This learning by observation occurred irrespective of the object chosen by the demonstrators as the positive choice and was more rapid than the learning that occurred during the conditioning of animals. The task was performed correctly without significant errors and further conditioning for 5 days. These results show that observational learning can occur in invertebrates.

Problem solving ability of Octopus vulgaris Lamarck (Mollusca, Cephalopoda).

Experiments presented in this study show that Octopus vulgaris Lamarck is able to open transparent glass jars closed with a plastic plug and containing a live crab (Carcinus mediterraneus). The animals remove the plus (Operandum: O) and seize the crab (Predation :P) in one single attack. The number of unsuccessful attacks appears to decrease over a series of trials (p less than .01); during the same period exploration time remains unchanged. There is a statistically significant increase in performance over trials for O (p less than .01) and P (p less than .05) mean times analyzed by single factor ANOVA, suggesting that the learning process is accomplished either by stimulus-response association or by trial and error. We propose that Octopus vulgaris is capable of learning the solutions of both problems, Operandum and Predation, thus showing a highly developed ability of "integration" of the behavioral program.

Is there "pain" in Invertebrates?

In contrast to nociception, the perception of pain, or pain experience, remains a subjective notion applicable to humans, but untestable with animals. Yet, when defined operationally as a physiological response induced in an animal by stimuli painful to humans, and resulting in a protective stimulus avoidance response, pain is amenable to testing with non-human subjects. This paper considers a series of examples showing responses to stimuli that are both painful (nociceptive) and responsible for eliciting natural self-preserving behavior in Invertebrates. Consideration is also given to the evolution and possible mechanism underlying the "pain-system" in Invertebrates.