Whereas standard stair descent was most demanding on the knee joint, by increasing riser height the greatest demand was placed on the ankle joint. According to dynamometrynormalized joint moments, at the point of highest demand the ankle and knee joints were operating at 89 and 103% of maximum capacity in stair descent, respectively. Conversely, descending increased rise stairs required the ankle joint to work at 175%, and the knee joint 76% of maximum moment capacity at the point of highest demand. As discussed above, the ankle operating beyond maximal capacities after training may be attributed to a gastrocnemius bi-articular action for (dynamic) stair descent, and uni-articular action for (static) isokinetic dynamometry testing.

No falls or events of postural instability occurred in stair descent in our cohort, indicating that these community-dwelling older adults were capable of coping with excessive demand at the ankle joint. However, for adults with functional limitations at the ankle it would be extremely difficult to safely descend stairs of increased rise unaided. Where environmental aids are not present, older adults can adopt non-cyclical gait strategies (e.g., step-by-step or side-stepping) to control gait speed, without prolonging single-leg support in stair descent. For example, sidestepping can be used to reduce ankle plantarflexor and hip extensor demand, without enlarging the lead limb joint moments upon step contact (King et al., 2018). These compensatory strategies, however, involve the preferential use of a single lead

TABLE 5 | Centre of pressure (CoP) and centre of mass (CoM) separation peaks of older adults when descending stairs of different dimensions before and after 16 weeks of exercise training.

Values are mean ± SD (n = 8). Significant difference: training ∗p < 0.05.

TABLE 6 | Velocity and acceleration of older adult's center of mass (CoM) when descending stairs of different dimensions before and after 16 weeks of exercise training.

Values are mean ± SD (n = 8). Significant difference: training ∗p < 0.05.

limb, which may be unsuitable to recover bilateral function in individuals with unilateral muscle (Metcalfe et al., 2013).

For standard stairs, exercise training was effective in alleviating the mechanical knee joint work in descent. From an interventional perspective, mechanical joint work was more heavily influenced by stair dimension, than exercise training. Increased rise stairs required greater negative work at the ankle (−0.97 J) and hip (−0.24 J) joints in descent, when compared to standard stairs (**Figure 2**). Riener et al. (2002) found similar involvement for increased joint power maximums at the ankle (67.3%) and hip (24.3%) when increasing stair inclination during descent. Stair descent involves large dorsiflexion joint range and support moments; if an individual lacks ankle joint range and/or functional strength when descending increased rise stairs, the lowering of mass to the below step will be greatly compromised. For this reason, our older cohort underwent lower limb resistance exercise, with rate of torque development and stretching specifically for the ankle plantarflexors, with which to enhance maximum dorsiflexion motion and support moments in the lowering phase. Thus, an important step in developing future exercise-based interventions would be to prioritize the ankle joint, particularly targeting plantarflexor muscular capacities for older adults in an ecologically valid environment.

The altered gait pattern required to safely descend increased rise stairs not only imposed greater functional demand, but also challenged movement control. Descending stairs of increased rise resulted in higher CoM and lead foot velocities, and CoM acceleration toward the ground, when lowering to the below step. The descriptive statistics indicate greater variability in CoM

motion (see **Table 6**) when descending increased rise stairs. This is reasonable considering the greater vertical distance that must be overcome when stepping downward with increased rise steps. It is equally plausible that this added demand also contributed to the slightly faster CoM peak downward acceleration (from −3.7 m/s2 to −4.4 m/s2 ; d = 0.28) in the control group when descending increased rise stairs, after 16 weeks of no training. The speed at which CoM downward velocity is arrested on landing in stair descent is mediated by the ankle joint's capability to produce eccentric torque at high angular velocities (Buckley et al., 2013). With older adults already operating at near maximum ankle capacity for standard stairs, and in-excess of maximum for increased rise stairs, individuals would have to manipulate their movement strategy. This was characterized by a reduced CoP-CoM displacement in the frontal plane (i.e., medial-lateral direction), and larger CoP-CoM displacement in the sagittal plane (i.e., anterior-posterior) during increased rise stair descent.

Descending increased rise steps (255 mm) requires greater plantarflexor and dorsiflexor activity upon landing (Gerstle et al., 2018), which may reflect a challenge to medial-lateral stability. Increased maximal ankle capacities following exercise training may have contributed to assist in controlling medial-lateral CoM stability. Progressing forward into single-leg support with the right foot leaving the above, increased rise step, the CoM was positioned further behind the CoP of the left foot, indicating greater posterior lean in descent (**Figures 3A,C**). Thereafter, from the right swing phase into double support the CoM was positioned closer to the CoP, indicating reduced anterior lean. No previous study has reported functional demand and balance control at the ankle, knee, and hip joints during increased rise stair descent for healthy, older adults. Greater trunk posterior leaning when lowering body mass further on to the below step, and lesser anterior leaning, upon the step contact (and mass acceptance) can be seen as a safe strategy in preventing uncontrolled CoM acceleration.

### Effects of Exercise Training at Increased Riser Height

Older adults adopt cautious movement strategies in stair descent to safely maintain lowering of mass, including prolonging the trail limb muscle co-contraction at the ankle and knee (Buckley et al., 2013) and reducing lead foot heel clearance (Kunzler et al., 2017). By increasing the riser height, and therefore task demand, our healthy older adults adopted alternative movement strategies to control safe stair descent. However, 16 weeks of exercise training was ineffective in enabling participants to further cope with the additional demands of increased rise stair descent. Our increased riser steps (255 mm), were only 15% higher than the maximum rise recommended for new private domestic stairs (220 mm) (HM Government, 2013). Communitydwelling older adults can be expected to negotiate similar riser heights in private and public buildings constructed prior to new national regulations.

The knee joint produced greater moment in single-leg support on increased rise stairs after exercise training. However, when normalized to dynamometry-assessed maximum eccentric moments, the exercise training had no effect on lowerlimb joint moments in stair descent. Similarly, the amount of negative work performed by a lower-limb joint was constrained by stair rise, and not influenced by exercise training. Training improvements in ankle torque production may have been sufficient to support the contralateral limb, as body mass was lowered to the step below, but for increased rise stairs, it appears ankle adaptations could not overcome the additional demand of lowering greater vertical distance. In this case, greater knee extension moments when descending increased rise stairs following training may have compensated, particularly to counter large knee flexion angles, which exceed those of standard rise stairs (Protopapadaki et al., 2007).

In the present study, participants descended each stair step-over-step, at a self-selected pace. Therefore, by necessity the descent tasks were different between stair configurations, but theoretically identical before and after training. However, as stride frequency lowered for increased rise stair descent after training, the same motor task was not being performed. Whether this reflects a more stable and controlled gait is unclear as movement control remained unchanged after exercise. Unsurprisingly, exercise training did not affect balance control (i.e., CoP-CoM separation) when descending increased rise stairs. The greater downward distance to the below step required not only an altered movement strategy to control CoM velocity, but also the ankle joint to operate beyond functional capacity. The increased rise was too high to "allow" any adaptation after exercise training. This is also supported by the downward CoM velocity and acceleration, which remained unchanged. The improvements in maximum functional capabilities at the ankle, knee and hip were nullified by the requirement to further control CoM acceleration and to generate sufficient external joint moments with which to alter postural strategies (i.e., CoP-CoM separation). Our older cohort was instructed to descend stairs using the stepover-step gait pattern, and therefore participants could not adapt their strategy to reduce the biomechanical demand. This further indicates that the increased rise stairs posed maximal demand, for which exercise training could not overcome, as opposed to older adults having to adopt differential gait strategies (e.g., side-stepping) to cope with increase rise descent.

### CONCLUSION

This study demonstrated that combined, lower-limb resistance and stretching exercise training can confer functional improvements in older adults when descending a staircase with a standard 170 mm riser height. Specifically, enabling our older cohort to redistribute lower-limb joint moments and adopt movement strategies to cope with the task demands of stair descent, including a safer balance control strategy. However, exercise training could not overcome the extra biomechanical demands imposed by increasing riser height by 50%. The post-exercise improvements at 16 weeks in the standard riser descent task highlights the relevance of incorporating appropriate exercise training in interventions aiming at improving daily stair safety in older people.

### ETHICS STATEMENT

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This study was carried out in accordance with the recommendations of the Ethics and Governance guidelines of the Manchester Metropolitan University, with written informed consent from all participants. All participants provided written informed consent in accordance with the Declaration of Helsinki. The protocol was approved by the ethics committee of the Manchester Metropolitan University, United Kingdom. We can confirm no vulnerable populations were involved as participants in the study.

### REFERENCES

### AUTHOR CONTRIBUTIONS

NR, CM, JB, VB, DJ, and MR contributed to the conception and design of the study. NR performed the data collection. JG performed the subsequent data and statistical analyses. JG, NR, CM, JB, VB, and MR interpreted the results of the research. JG wrote the first draft of the manuscript. NR, CM, JB, VB, and MR reviewed the manuscript, as JG revised the subsequent manuscript versions. All authors contributed to the final manuscript revision, and read and approved the submitted version.

### FUNDING

This study was funded by the New Dynamics of Ageing (NDA) Programme (Grant: ES/G037310/1) who had no involvement with this study.

**Conflict of Interest Statement:** MR was employed by the company Building Research Establishment (BRE) during the research study. MR is currently employed by the company Rise and Going Consultancy. The BRE's goal is "to make the built environment better for all," and the company supports independent research by allowing researchers and engineers collaborate together in creating evidence-based solutions.

The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

Copyright © 2019 Gavin, Reeves, Jones, Roys, Buckley, Baltzopoulos and Maganaris. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

# Comparison of Muscle Function, Bone Mineral Density and Body Composition of Early Starting and Later Starting Older Masters Athletes

Jessica Piasecki1 , Alex Ireland2 , Mathew Piasecki3 , Kevin Deere4 , Kimberley Hannam4 , Jonathan Tobias4 and Jamie S. McPhee2,5 \*

1 Sport, Health and Performance Enhancement Research Centre, Nottingham Trent University, Nottingham, United Kingdom, 2 Department of Sport and Exercise Sciences, Musculoskeletal Science and Sports Medicine, Manchester Metropolitan University, Manchester, United Kingdom, 3 Clinical, Metabolic and Molecular Physiology, MRC-ARUK Centre for Musculoskeletal Ageing Research, National Institute for Health Research (NIHR) Nottingham Biomedical Research Centre, University of Nottingham, Nottingham, United Kingdom, 4 Musculoskeletal Research Unit, Bristol Medical School, University of Bristol, Bristol, United Kingdom, 5 Department of Physiology, University of Padua, Padua, Italy

#### Edited by:

Martin Burtscher, University of Innsbruck, Austria

#### Reviewed by:

Javier Brazo-Sayavera, Universidad de la República, Uruguay Rafael Timon, University of Extremadura, Spain

> \*Correspondence: Jamie S. McPhee<EMAIL_ADDRESS>
#### Specialty section:

This article was submitted to Exercise Physiology, a section of the journal Frontiers in Physiology

Received: 17 December 2018 Accepted: 30 July 2019 Published: 27 August 2019

#### Citation:

Piasecki J, Ireland A, Piasecki M, Deere K, Hannam K, Tobias J and McPhee JS (2019) Comparison of Muscle Function, Bone Mineral Density and Body Composition of Early Starting and Later Starting Older Masters Athletes. Front. Physiol. 10:1050. doi: 10.3389/fphys.2019.01050 Masters endurance runners can epitomize healthy aging; being reflective of the physiological processes of aging without the compounded effects of inactivity. The primary aim of the present study was to determine, using cross-sectional data, whether individuals taking up training after the age of 50 years can achieve the same level of athletic performance and musculoskeletal characteristics in their older age as those who trained all of their adult lives. A total of 150 master endurance runners [age 68 (5) years; 111 male, 39 female] were divided into early starters (training all of their adulthood) and late starters (started training after age 50 years). A comparative non-athletic group of 59 healthy older adults [age 73 (4) years; 30 female, 29 male] were additionally included for analysis. Training intensity, age-graded performance (AGP) and musculoskeletal assessments were performed. Results showed that there was no difference between athlete groups for training intensity or age-graded performance, despite the 30-year difference in training history. Body fat percentage and leg lean mass did not differ between athlete groups, but were 17% lower and 12% greater, respectively, in athlete groups compared with controls. Power normalized to body mass did not differ between any groups. Spine BMD was lower in late starters than controls, while early starters did not differ from late starters or controls. Hip BMD did not differ between any of the groups. These findings show that the Masters athletes we studied that started intense endurance running after the age of 50 years had lower body fat and higher leg lean mass compared to non-athletes. Body composition and athletic performance of the late starters was very similar to those who trained all of their adult lives.

Keywords: masters athletes, body composition, bone mineral density, starting age, endurance running

## INTRODUCTION

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There are growing numbers of older people training and competing as Masters runners in mass-participation endurance events, such as 5, 10 km, Half Marathon or full Marathons (Stevinson and Hickson, 2014; Lepers and Stapley, 2016). At the highest competitive level, Masters runners train 4–7 times per week at high intensity (Piasecki J. et al., 2018). Their overall lifestyle includes more physical activity than the general population, as they accumulate around 3-fold more low impacts as a result of voluntary movement during general activities, 20-fold more medium impacts and 200-fold more high impacts (measured in units of gravity, g) compared with typical older adults from the general population (Deere et al., 2016; Hannam et al., 2016b).

Intense training sustained through middle and into older age helps to maintain low body fat percentage (Piasecki et al., 2019) and, possibly, greater lean mass and skeletal muscle strength into old age. However, the available evidence regarding muscle mass and function of older Masters runners is conflicting. A recent meta-analysis highlighted research gaps due to most studies having low participant numbers with very few female athletes, limited outcome assessments and unclear demographics (Mckendry et al., 2018). For example, two studies (Power et al., 2012; Stenroth et al., 2016) showed higher maximal force or power in Masters endurance runners compared with non-athletic old, but several others showed no difference between Masters endurance runners and controls (Aagaard et al., 2007; Michaelis et al., 2008; Mikkelsen et al., 2013; Couppe et al., 2014; Piasecki et al., 2016; Piasecki J. et al., 2018) or even lower (Sipilä and Suominen, 1991; Sanada et al., 2009) maximal force or power in Masters endurance runners compared with non-athletic old. More recently, Fien et al. (2017) reported high levels of physical function, low body fat, and good health status of Masters athletes, but these characteristics declined with advancing age and because no comparison was made with non-athletic old it is not possible to know if these characteristics were any better than typical healthy old. Differences between studies in the athletic ranking of Masters athletes (e.g., highest achievements), age and years of training are possible explanations for the conflicting reports, but there is very little information available to show how the starting age influences physical performance, muscle mass, and body composition in later life.

In the general population, leg skeletal muscle mass declines progressively from age 30 years at a rate of approximately 8– 10% per decade (Lynch et al., 1999; Janssen et al., 2000; Lee et al., 2004; Mitchell et al., 2012; Moore et al., 2014) and maximal force and power decline progressively at approximately 10–12% per decade (Michaelis et al., 2008; Dodds et al., 2014; Bagley et al., 2019). Therefore, by age 50 years non-athletic men and women have typically undergone two decades of progressive muscle declines. If regular endurance running preserves muscle structure and function into older age (Ringsberg et al., 2001; Landi et al., 2018; Shiroma et al., 2018), then life-long athletes should have avoided these age- and lifestyle-dependent declines and should have greater muscle mass and function than those taking up running after the age of 50 years. Although no research is available to support these possible benefits of longer-term training, there is evidence to suggest that starting age is an important factor. A previous study of Masters tennis players found that the upper limb (radius, ulna, and humerus) bone circumference and bone mineral density were greater in those who started training at an early age compared with those starting at an older age (Ireland et al., 2014). However, this study did not examine muscles or bones of the legs that would be exposed to "impacts" during running. More crucially, the positive influence of playing tennis from a young age for arm bones conflicts with other data suggesting that long-term endurance running does not give higher bone mineral density or may even be detrimental for spine, leg or hip bone mineral density (MacKelvie et al., 2000; Wilks et al., 2009a; Piasecki J. et al., 2018).

There is a gap in available knowledge of whether the age at which people take up competitive endurance running influences musculoskeletal structure and function as well as body composition in later life. These questions are important because regular exercise is recommended as the best way to combat agerelated declines of the musculoskeletal system (Acsm, 1998; Kusy and Zielinski, 2015; Chief Medical Officer's Guidelines, 2018; Cruz-Jentoft et al., 2018) and the age at onset of exercise may affect health and performance-related indicators in later life. We therefore aimed to compare body composition, lean mass, maximal power, and hip and spine bone mineral density between Masters endurance runners competing all of their adult lives with those taking up competitive running after the age of 50 years. For reference, a non-athletic healthy older group was also included. It was hypothesized that athletic performance, lean mass, muscle power, and bone mineral density would be greater in trained compared with untrained adults, and greater in those with the longer history of athletic training compared to those starting athletic training after the age of 50 years.

### MATERIALS AND METHODS

### Study Design

The study was approved by the Regional Ethics Committee (North West England: 14/NW0275) as well the Local Ethics Committee of Manchester Metropolitan University. Written informed consent was obtained from all participants. Masters runners were recruited as part of the United Kingdom-based "VIBE" study (Deere et al., 2016; Hannam et al., 2016b). Male and female athletes were recruited from regional athletics meetings from across the United Kingdom. To be included, they must have been aged ≥59 years at the point of enrollment, had competed in endurance running in the past 12 months at a regional level and free from serious injury for over 2 years. They were invited to attend the research facility at Manchester Metropolitan University (United Kingdom) and were required to avoid training or competing for at least 48 h prior to attendance. The full cohort included 188 Masters Athletes, of which 33 were sprinters (events less than or equal to 400 m) and 150 were endurance runners (events greater than or equal to 800 m). Given the low number of sprinters (when separated to early starters, late starters, men, and women) and the known differences

in muscle mass and bone mineral density between sprinters and endurance runners (Gast et al., 2013; Kusy and Zielinski, 2015; Piasecki J. et al., 2018), the sprinters were excluded from further analysis for this particular study. Thus, a total of 150 Masters endurance runners were available for analysis. Mean age-graded performance (AGP) was determined by taking the athlete's highest ranked performance within the last 2 years and expressing it as a percentage of the world record for that age and distance. AGP ranged from 77–92% across the cohort, indicating a very high level of performance relative to age group records. For example, a 3 h and 30 min marathon for a 70-year-old man gives an age-graded performance of 80%, as described previously (Piasecki J. et al., 2018).

Data for control participants was taken from the MYOAGE study, a European multi-center study of healthy aging and the methodology has been described previously (McPhee et al., 2013). These participants were recruited by focused advertisement in newspapers, the third-generation university and the association of emeriti and universities, thus selecting cognitively active individuals living independently and without mobility impairments. An initial telephone interview was used to screen volunteers, which asked for self-reported number of exercise and social activity sessions per week over the past 3 years. Those who were sedentary defined as not involved in any regular activity sessions, as well as those involved in any form of regular, intense athletic or gym training were excluded from the control group. The controls did not complete accelerometrybased activity monitoring for use in this study. For the purpose of this study the 59 older participants recruited at the Manchester, United Kingdom, site and with complete data including hip and spine bone mineral density measurements were used.

Therefore, the total number of complete datasets available for analysis was 209 and all participants completed measurements at the same laboratory and using the same equipment. Of those, 140 were male and 69 were female. For this study, the athletes were divided into early starter (ES) and late starter (LS) athletes. These groups were based on answers from a questionnaire asking each athlete to self-report the number of hours spent training (0–1, 2– 3, 4–7, over 7 per week) during different stages of their adult life (18–29, 30–49 and 50 years and over). Early starter athletes were defined as those that had taken part in competitive endurance running throughout their adult lives, reporting intense training and competition at ages 18–29, 30–49, and 50 years and over. Late starter athletes were defined as those that had taken up intense training and competition after the age of 50 years, with no previous competitive training history. See **Table 1** for participant characteristics.

### Questionnaires

Participants provided demographic, general health, lifestyle, and physical activity information by questionnaire [described previously (Hannam et al., 2016b)]. Self-rated health was reported on a scale from very good through to very poor, along with details of any diseases or prescribed medications. Current and history of smoking was collected, including number of cigarettes smoked per normal day, age started smoking and the duration, if ever, of smoking. Typical alcohol consumption was recorded and the type of alcohol. Highest level of education was also recorded. Questionnaire response data has been provided in **Supplementary Table S1**.

### Accelerometry

Accelerometry data was collected only in the athletes and has been described previously (Deere et al., 2016; Piasecki J. et al., 2018). Each athlete received a GCDC × 16–1c (Gulf Coast Data Concepts, Waveland, MS, United States), which was placed in a Velcro strap and worn tightly around the waist with the accelerometer device placed over their right hip. Each athlete wore a monitor for seven consecutive days, only removing it when showering, bathing, swimming, and sleeping. Each athlete completed a time sheet over the 7-day period to record when the monitor was worn and to indicate any reason why that day was not representative of their usual routine. Accelerometers were configured with standardized settings prior to participant use with a sampling frequency of 50 Hz, a deadband setting of 0.1 g (the threshold which must be exceeded before a recording is made) and a timeout setting of 10 s (meaning that a single sample every 10 s is taken even if the recording is <0.1 g) (Deere et al., 2016). Once the 7 day period of use was completed the participant returned the accelerometer to the research facility by post. The raw accelerometry data was uploaded to a secure shared drive and read into Stata 13 (StataCorp, College Station, TX, United States). A standardized cleaning and processing procedure was used and has been described in detail previously (Deere et al., 2016). In short, the Y-axis accelerations dataset was cleaned to remove any movement artifacts and any periods of nil data collection, most likely due to the participant not wearing the accelerometer. Activity data were normalized based on seven valid days of 14 h with ≥10 h recording time. Y-axis peaks were calculated based on accelerations that were higher than the previous and subsequent reading and recorded within 14 pre-specified g bands. These were condensed to three impact bands; low (≥0.5 to <1.0 g), medium (≥1.0 to <1.5 g) and higher (≥1.5 g) impact. All g values represent g over and above 1 g from earth's gravitational force (Hannam et al., 2016a).

### DXA Scans

Standing height was measured to the nearest millimeter and body mass was measured to the nearest 0.1 kg. Whole body, total hip and lumbar spine dual energy X-ray absorptiometry (DXA: Lunar Prodigy Advanced, GE Healthcare, encore version 10.50.086, London, United Kingdom) scans were performed while the participant lay supine wearing a light cotton t-shirt to reduce measurement errors due to clothing absorption. Body composition (fat mass and lean mass) was taken from results of total body scans and regional analysis of legs and arms. Bone mineral density (BMD, g.cm−2 ) was taken from hip and spine scans. All measurements were recorded after manual adjustment of the regions of interest. Repeat total body and hip scans were performed in eight participants within 1 month of the first scan. Using these repeat scans, the short-term error for our laboratory was 2.0% for hip BMD, 0.9% for spine BMD and 0.01% for whole body lean mass.

Body Composition of Masters Athletes

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#### TABLE 1 | Participant characteristics.

Data are mean ± SD. Age and sex were included as covariates, except where indicated by awhere sex was the only covariate. bData are presented as median (25th/75th) quartiles. 95% Confidence intervals (95%CI). Bold text is used to highlight statistically significant difference (p<0.05).

### Muscle Function

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The investigators provided verbal instructions and a physical demonstration of the muscle function tests. Participants were allowed one practice immediately before the actual assessed trials, which acted as a specific warm up and also confirmed that the instructions were understood. In all cases, the muscle function tests were completed between 10 am and 3 pm.

Hand grip strength was measured using the Jamar dynamometer handle (Sammons Preston Inc., Bolingbrook, IL, United States) as previously described (Hannam et al., 2017). The width of the dynamometer was adjusted for each participant separately. Participants were instructed to stand upright with the arm fully extended along the body, maintaining approximately 5 cm gap between the wrist and the hip or upper leg (so that the hand was not rested against the body). Participants were instructed to squeeze against the handle as hard possible for 3 seconds. Grip strength was measured three times and recorded in kilograms to the nearest 0.1 kg. For the purpose of this study, the best of three attempts was included in further analysis.

A Leonardo Jump Mechanography Platform (Leonardo Software version 4.2: Novotiec Medical GmbH, Pforzheim, Germany) was used to assess lower limb muscle power during a countermovement vertical jump, as described previously (Hannam et al., 2017). Results for both absolute (W) and relative (W/kg) power were recorded. Briefly, a two-footed countermovement jump was performed starting with feet approximately 30 cm apart (slightly narrower than shoulder width) and standing upright on the force plates. Force was sampled at 800 Hz. Participants flexed at the knees before extending as forcefully as possible to take off for the jump. Jumps were performed with a trained research assistant in close proximity to intervene in case of a trip or fall. Each participant repeated the jump sequence three times, with approximately 60 seconds rest between efforts. The jump with the highest value for power was used for statistical analysis.

### Statistical Analysis

Statistical analysis was performed using SPSSv21 (IBM, United States). Normality of distribution was assessed using the Shapiro–Wilk test, which showed that all data was normally distributed (presented as mean ± standard deviation) except for accelerometry data (presented as median (25th/75th) quartiles). Two-way ANOVAs (three group and two sex; all p > 0.05) showed no group × sex interactions for any of the variables so for this reason, together with the relatively small numbers of females in the ES group, data from both sexes were included within the same analysis. Univariate ANOVA (ANCOVA) analysis was used to identify differences between the three groups (ES, LS, and C) with age and sex added as covariates, to account for the different mean ages and proportions of males and females between groups. Where significant differences were found, Fisher's Least Significant Difference pairwise comparisons were performed. Differences between groups were considered statistically significant at p < 0.05.

As participants included in our study were initially recruited to address other primary research questions, sample sizes for our analyses were fixed by the data already available. Our power to determine small, medium and large effect sizes can be ascertained from the observed effect sizes for the different outcome variables based on the partial eta squared (η 2p), as recommended by Lakens (2013) and O'Keefe (2007) for use with ANOVA analysis including covariates (age and sex). A small effect size was set at η 2p = 0.01, a medium effect size was set at η 2p = 0.06 and a large effect size was set at η 2p = 0.14. The values provided in the Results section show that the study was sufficiently powered to detect even small differences between groups for the main outcome variables of body fat percentage, lean mass, spine BMD, and vertical jump power.

## RESULTS

### Participant Characteristics

Participant characteristics are shown in **Table 1** and other health and demographic information has been provided in **Supplementary Table S1**. The ES athletes had been training for 30 years longer than LS athletes (p < 0.0005). Pairwise comparison showed there was no significant difference between ES and LS for AGP (p = 0.294). Accelerometry collected over 7 days showed similar impacts in the low, medium, and high g bands between ES and LS groups (p = 0.089, 0.799, and 0.352, respectively, as shown in **Table 1**).

There was no significant difference between groups for height. However, age, body mass, and BMI were significantly different between groups (all p < 0.0005). Pairwise comparisons showed that C were older than ES and LS, with no difference between ES and LS groups. Body mass and BMI were similar for the ES and LS athletes, but significantly greater for C compared with ES and LS groups (both p < 0.0005). The main reason for greater body mass in C was the higher body fat percentage (**Figure 1B**).

### Muscle Function and Body Composition

Total body fat percentage differed significantly between groups (p < 0.0005; η 2p = 0.380) due to approximately 17% lower values for both athlete groups than C (p < 0.01; **Figure 1A**). Leg lean mass differed significantly between groups (p < 0.0005; η 2p = 0.098; **Figure 1B**), being approximately 12% greater in both athlete groups than C (p < 0.003). Appendicular lean mass was also different between groups (p = 0.003; η 2p = 0.062) and although values were similar between athlete groups, the LS had a significantly greater lean mass than C (p = 0.001). Total body lean mass did not differ significantly between groups (η 2p = 0.028; **Table 2**).

Spine BMD (p = 0.004; η 2p = 0.053), BMC (p = 0.001; η 2p = 0.064), and Area (p = 0.014; η 2p = 0.041) were significantly different between groups. There were no differences for these measurements between athlete groups, but LS had lower BMD (p = 0.001; η 2p = 0.053), BMC (p < 0.0005; η 2p = 0.063), and Area (p = 0.004; η 2p = 0.041) compared to C. There was no significant difference in Hip BMD (η 2p = 0.024), BMC (η 2p = 0.020) or Area (η 2p = 0.005) between any of the groups (**Table 2**).

Maximal grip strength (η 2p = 0.020) and vertical jump power relative to body mass (η 2p = 0.005) did not differ

significantly between groups (**Table 2**). However, vertical jump power in absolute values was different between groups (p = 0.020; η 2p = 0.017); values were similar between athlete groups, but controls exhibited a greater power than LS (p = 0.006).

### DISCUSSION

The novel contribution of the present study is the focus on the starting age of Masters endurance runners in relation to later life performance and musculoskeletal health indicators. This is supported by the relatively large sample size, depth of physiological profiling and, for the Masters athletes, objective characterization of habitual physical activities. Our findings demonstrate that the men and women of the LS group with no previous history of intense training or competition before the age of 50 years, had by the age of 70 years very similar training intensity, athletic performance, body fat percentage and leg lean mass to athletes of the ES group whom had accumulated 30 extra years of training and competition (**Table 2** and **Figure 1**). Both athlete groups had lower body fat and greater leg lean mass than healthy non-athletic controls, but spine BMD was lower in LS than in C. These findings suggest that starting regular, intense endurance running at, or after, the age of 50 years is not too late to compete at the highest level in Masters endurance running or to significantly delay accumulation of body fat and loss of leg lean mass in older age.

The physical activity levels of ES and LS equates to around 3 fold greater level of low impacts, 20-fold greater level of medium impacts and 200-fold greater level of high impacts per week compared with reference values that were previously reported for older adults from the general population (Hannam et al., 2016b). Past studies have demonstrated both a dose-response and an intensity-dependent response to regular exercise training, as more frequent and higher intensity training confer beneficial health- and performance- adaptations (Swain and Franklin, 2006; Bruce et al., 2008; McPhee et al., 2016). The high impacts and activity levels due to more vigorous intensity movements alongside the associated energy expenditure is the most likely reason why both athlete groups avoided the usual increase in adiposity with advancing older age (Elhakeem et al., 2017).

Appendicular and leg lean mass were similar between athlete groups, but both athlete groups had higher leg lean mass than C (**Table 2** and **Figure 1**). This is in line with two studies (Mikkelsen et al., 2013; Couppe et al., 2014) that reported greater muscle size of Masters endurance athletes than age-matched controls. These past studies (Mikkelsen et al., 2013; Couppe et al., 2014) included only 15 Masters athletes, compared with our much larger sample of 150 endurance runners with detailed activity tracking by accelerometry and confirmation of the high AGP. The prevailing view is that resistance exercise is needed to reduce risk of sarcopenia in old age (Cruz-Jentoft et al., 2018; Lee et al., 2018; Vlietstra et al., 2018). Our results suggest that long term intense endurance running is also effective, and that it does not matter if this activity is taken up after the age of 50 years. However, from the available data we are not able to identify an older age at which benefits of intense endurance running are diminished compared with those training for all of their adult lives.

One of the benefits of having larger muscles is the potential to develop greater muscle force and power. In our study, power measured by vertical jump was not different between athlete groups, but actually lower for LS compared with C. However, this difference between groups disappeared when normalized to total body mass (**Table 2**). Previous studies also reported similar vertical jump power in Masters endurance runners and agematched non-athletic individuals (Michaelis et al., 2008). We did not determine the reasons why the larger muscles of endurance runners do not produce greater power, but it is likely related to the characteristic "slow" muscle fiber contractile properties of endurance athletes (Tanaka and Seals, 2008) which makes energy turnover more economical, but gives lower power as the product

#### TABLE 2 | Musculoskeletal characteristics.

Data are presented as mean ± SD. Age and sex were included as covariates. Values highlighted in bold text identify significant differences between groups. BMD, Bone mineral density; BMC, Bone mineral content; %Female as displayed inTable 1. 95%Confidence intervals (95%CI).

fphys-10-01050 August 23, 2019 Time: 18:22 # 7

Piasecki et al.

of force x velocity of contractions (Michaelis et al., 2008). We did not measure knee extensor maximal force, but the available evidence is conflicting over the possibility that Master endurance runners have greater maximal force, mainly due to heterogeneity in study populations and several studies with low sample size (Mckendry et al., 2018).

The LS athletes had lower spine BMD, BMC, and Area than C, raising the possibility that starting regular intense endurance running after age 50 years may be detrimental to spine bone health. This may seem counter-intuitive, as it is proposed that regular exercise with high impacts can improve bone mineral density and bone strength (Ireland et al., 2011, 2013) as high muscular forces stimulate osteogenic responses (Frost, 1987a,b). However, this previous literature is primarily based on observations of long limb bones (Wilks et al., 2009a,b; Ireland et al., 2011, 2014). For example, previous studies showed that the limb bone circumference of Masters athletes (33–94 years old) was greater than that of age-matched sedentary controls (Wilks et al., 2009b). These beneficial effects may be limited to limb bones, younger ages or participation in sprint or power activities (Warden et al., 2007, 2014; Wilks et al., 2009b; Ireland et al., 2014). Previous research has also demonstrated an inverse association between the amount of low-impact physical activity and hip and spine BMD (Johansson et al., 2015; Hannam et al., 2016b; Piasecki J. et al., 2018). Although the explanation is lacking for why completing lots of low impact activity may be detrimental for bones, it is well known that bone responses depend on the type of physical activity being performed (Nichols et al., 2003; Velez et al., 2008; Wilks et al., 2009a; Piasecki J. et al., 2018; Pollock et al., 2018). For instance, sprinting is associated with greater hip, spine and tibial BMD compared to endurance running or non-athlete controls (Wilks et al., 2009a; Piasecki J. et al., 2018). Although not measured in the present study, Vitamin D and calcium intake are also associated with skeletal health in older age (Cashman, 2007). In particular, Vitamin D deficiency and low calcium intake can lead to low bone mineral density and they are used as supplements to combat osteoporosis (Grados et al., 2003a,b). Therefore, any interventions later in life to improve bone health may need to include nutritional supplementation, and also consider sprint or jumping activities in addition to regular prolonged endurance running.

Overall, our findings build upon available evidence that shortterm exercise can improve some features of musculoskeletal health in middle- and older-age (McPhee et al., 2016) and have highlighted possible benefits of very long-term exercise (Velez et al., 2008; Wilks et al., 2009a,b; Nowak et al., 2010; Ireland et al., 2015; Mckendry et al., 2018), although these benefits may not be present for lumbar spine bones. The novel contributions of the present study describing the very long-term intense training and the focus on starting age are important because starting at a later age risks the possibility that irreversible age-related declines have already occurred and would limit adaptations that improve health and performance. For example, age-related reduction of maximal heart rate limits cardiac output and therefore peak aerobic capacity. Within the musculoskeletal system the skeletal muscle mass, strength and power decline from the fourth decade of life (Lynch et al., 1999; Janssen et al., 2000; Silva et al., 2010; Reid and Fielding, 2012; Moore et al., 2014; Pantoja et al., 2016; Fien et al., 2017; Bagley et al., 2019). The extent to which these changes are related to irreversible reductions of muscle fiber numbers (Lexell et al., 1988; McPhee et al., 2018) and motor units (Tomlinson and Irving, 1977; Piasecki M. et al., 2018; Piasecki et al., 2019) remains unknown. In this respect, the findings of the present study are encouraging. The implication based on observations made from the participants in our study, is that starting intense training in middle age, approximately 20 years after musculoskeletal declines are detectable in the general population, is not a disadvantage compared to training throughout adult life when it comes to maintaining leg muscle mass, preventing accumulation of excessive fat mass and for athletic performance in older age.

The findings of this study may inform practitioners when recommending physical activity for older adults aiming to reduce fat mass, gain leg lean mass or improve bone mineral density. The new information showing that long-term endurance training is associated with greater leg lean mass should be considered by policy makers as an alternative, or addition, to resistance exercise to combat sarcopenia. One note of caution, however, is that the lower bone mineral density of the spine in LS may increase the risk of late life bone injuries.

### Study Limitations

The grouping of ES and LS has relied on self-reports and the cross-sectional study design prevents any causal relationships from being established. Our sample size of 209 provided sufficient power to detect even small differences in means between comparison groups, but we cannot rule out the possibility that there may be very small differences in outcomes between the ES and LS groups which we were unable to detect. There is also a risk of differences in data handling and screening of bone results (e.g., for spondylosis) when comparing the results of the Masters athletes with those of the controls, where data were collected approximately 5 years apart. However, care was taken to follow exactly the same protocols as far as possible and the assessments were made in the same laboratory using the same equipment. The values for body composition and lean mass of the controls are within the ranges previously published for non-athletic but otherwise healthy older adults from larger, multi-center studies (Bijlsma et al., 2013; Bauer et al., 2015; Coulson et al., 2017; Verlaan et al., 2017). Nevertheless, interpretation of outcome comparisons for the control group is limited because they did not complete the same physical activity assessments as the Masters athletes. The athletes were recruited based on their athletic performance over the previous 2 years. Within this time period there may have been fluctuations in activity levels due to injury or illness which were not captured within this study. Future studies could shorten the time period taken into account when recruiting the athletes or more carefully consider their time spent inactive. More detailed information about youth physical activity may also add further insights. A B-PAQ questionnaire (Weeks and Beck, 2008) may be used in future for this purpose. Studies of Masters athletes carry a possible bias because any individuals developing injury or disease may cease competing and would not be available

for recruitment. Masters athletes tend to be better educated, of higher socio-economic status and with fewer diseases than the general population (**Supplementary Table S1**), so the results of such studies may not be generalized to the wider population. Finally, we cannot rule out possible differences in energy intake or nutritional status, particularly of Vitamin D and calcium that influence musculoskeletal health, between athletes and controls because this was not measured in our study.

### CONCLUSION

The Masters athletes within our sample taking up intense endurance running after the age of 50 years had lower body fat and higher leg lean mass than non-athletes by the age of 70 years and the values for body composition and athletic performance of the late starters were very similar to those of people whom had trained all of their adult lives.

### ETHICS STATEMENT

The study was conducted in accordance with the Declaration of Helsinki and approved by the University Research Ethics

### REFERENCES

Committee and the National Research Ethics Committee Northwest (14/NW0275). All participants provided written informed consent.

### AUTHOR CONTRIBUTIONS

All authors contributed to the data collection, analyses, and wrote the manuscript. JP led the write up and directed the analysis of data.

### FUNDING

This project was supported by the funding from European Union FP7 ("MYOAGE," #223576) and Medical Research Council (MR/K024973/1).

### SUPPLEMENTARY MATERIAL

The Supplementary Material for this article can be found online at: https://www.frontiersin.org/articles/10.3389/fphys. 2019.01050/full#supplementary-material

and mechanical stress in connective tissue. Age 36:9665. doi: 10.1007/s11357- 014-9665-9

**Conflict of Interest Statement:** The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

Copyright © 2019 Piasecki, Ireland, Piasecki, Deere, Hannam, Tobias and McPhee. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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**7. Management**

*Invasive Species - Introduction Pathways, Economic Impact, and Possible Management Options*

**6. Invasive potential and global distribution**

by three basic components:

**Figure 5.**

According to Begon et al. [25], any species distribution is limited and governed

1.The ability of the species to reach a potential site (introduction pathway)

*Tunnels in ripe tomato fruits excavated by the larvae of Tuta absoluta (photos: Antonio Biondi).*

2.Capacity to develop in specific environmental conditions (establishment)

TLM is a highly invasive insect pest of tomato crop [1, 6]. The moth was first reported in Europe (Eastern Spain) in 2006 [19]. The introduction in Spain is believed to be from a single population in Chile [26]. Three years later, it was

reported in Turkey, the fourth largest producer of tomato in the world, in 2009 [27]. It spread then across Europe and North African countries [6, 28] and Asian countries [23]. According to Seebens et al. [29], most of the invasion occurred during the last 40 years due to increased globalization and trading among continents. The possible introduction pathways for *T. absoluta* include tomato fruits, packing materials of tomato, eggplant and pepper, and planting material [30]. Santana et al. [31] studied the global geographic distribution of TLM using a combination of spatial distribution models as well as the current distribution of the pest. They showed that the suitable areas for *T. absoluta* include North and Central Americas, Africa, Europe, Asia, and Oceania at present time and in the future. Additionally, their model showed that large tomato-producing countries such as China, USA, and Mexico, where the moth

3.The ability to compete with other species occupying the same habitat

**82**

Because of its high invasiveness and economic significance, management of *T. absoluta* could be carried out at local, regional, and international levels. The management can be divided into pre-invasion and post-invasion measures. The former are mainly preventive including strict quarantine measures, inspection of tomato consignments, and treatment, when necessary, with proper fumigants before shipping. Endorsement, by countries, of *T. absoluta* as pest of a high risk in quarantine list is essential. *T. absoluta* population management in invaded countries could significantly lower the invasion risk to neighboring non-invaded ones [23, 35]. Establishment of regional network to connect research entomologists, policy-makers, and major stakeholders from all invaded as well as threatened countries [23]. Such network and platforms are supposed to coordinate joint research activities and validation of newly developed management technologies before being applied in the field.

The post-invasion management of *T. absoluta* is to try to eradicate the pest at an early stage of invasion if possible, otherwise a sustainable containment strategy based on integrated pest management is recommended.

In native and invaded areas in the world, current IPM components against TLM include the following:


**3.2. NAC family**

The NAM/ATAF/CUC (NAC) TFs contain a unique feature, a conserved N-terminus DBD and a dissimilar C-terminus regulatory domain, and they are spread across the plant kingdom [37]. The name NAC is an abbreviation of three genes designated as no apical meristem (*NAM*), *Arabidopsis* transcription activation factor (*ATAF*), and cup-shaped cotyledon (*CUC*), which encode proteins containing homologous sequences as the NAC domain [49, 50]. *NAM* isolated from petunia by Souer et al. [49] was the first gene demonstrated to encode a NAC protein, followed by the gene *CUC2* from *Arabidopsis* [50]. Mutation of both genes resulted in the absence of apical shoot meristems [51] and led to floral abnormalities such as the alteration of petal primordia positions during the development stage. These evidences show that the TFs NAM and CUC play important roles in shoot apical meristem formation and determine the organ primordia positions in the floral meristem [49, 50]. But in relation to abiotic stress, the wheat genes *TaNAC2* [52] and *TaNAC69* [53] were strongly expressed under water deficiency and salinity. TaNAC67 was found to decrease the cell membrane instability, preventing water loss and enhancing other physiological processes that were considered to be responsive to drought, low temperature, and salt stress [54]. In maize, ZmNAC052, Zma000584, Zma006493, Zma001259z [55], and ZmSNAC1 showed increased transcription levels under water defi‐ ciency, indicating their potential role in drought tolerance regulation. In rice, SNAC1 [56], OsNAC5 [57], –6 [58], –9 [59], and –10 [60] altered the root structure for plant adaptation during drought. Further, the overexpression of *OsNAC045* [19] and *OsNAC52* [61] induced ABA sensitivity and conferred drought resistance in transgenic rice and transgenic *Arabidopsis,* respectively.

#### **3.3. MYBs family**

MYB is a group of ancient TFs found in viruses [62] and eukaryotes such as plants, animals, and fungi [63]. The first gene (c1) identified to encode MYB in plants was from *Zea Mays* [64]. MYB TFs contain a conserved DBD called MYB domain characterized by one to three imperfect repeated amino acid sequences (R1, R2, and R3). Each repeat sequence has around 50–53 amino acids which form three α-helixes [65]. These three α-helices form a helix-turn-helix structure when interacting with DNA. The MYB TFs are mostly classified into three classes according to the number of the MYB domain repeats: R1-MYB, R2R3-MYB [66], and R1R2R3-MYB [67]. These TFs participate in responses to dehydration, salt, cold, and drought [20, 68, 69]. In maize, the ZmMYB-R1 was induced by ABA, drought, low temperature, high salt, and heat [70]. The overexpression of rice genes *OsMYB3R-2* [20], *OsMYB4* [71], and OsMYB48-1 [72] improved the adaptive response to drought and other stresses in transgenic plants. In wheat, TaMYB3R1 [73] and TaMYB30-B [7] were found to be potentially involved in drought adaptation.


**About the Editors**

#### **Paolo Biagi**

Paolo Biagi (Full Professor Ph.D.) is a Senior Researcher of Prehistory and Protohistory at Ca' Foscari University of Venice, Italy. He has carried out research and excavations in several countries of Europe, the Caucasian mountains of Georgia, the Indian Subcontinent, and the Arabian Peninsula. His main interests regard the archaeology of the coasts of the Arabian Sea and the Indus Valley, the exploitation of the highland zones, in particular the Alpine chain, the Pindus range, and the Caucasus, and the exploitation and circulation of raw material sources, chert and obsidian in particular.

#### **Elisabetta Starnini**

Elisabetta Starnini (Ph.D.) is an Associate Professor of Prehistory and Protohistory at the Department of Civilizations and Forms of Knowledge of the University of Pisa, Italy. She has researched and excavated in several European countries, including Italy, Hungary, and Greece, the Indian Subcontinent, and the Arabian Peninsula. Her main interests regard the scientific identification, exploitation, and circulation of raw materials, particularly rocks for prehistoric artifacts from the Palaeolithic to the Bronze Age, among which are chert, obsidian, and metamorphic rocks.