Within-individual Variance Research Articles

PSX-5 Repeatability of overreach, term and acuteness in Peruvian Paso horse

Abstract The Peruvian Paso Horse is considered one of the best saddle horses worldwide due to the gait that makes this horse perform one of the smoothest walks. The overreach, term, and acuteness are the most characteristic movements in the gait, making them the most desirable attributes in its breeding and reproduction. Repeatability of overreach, term, and acuteness in Peruvian Paso horses (PPH) have not been determined to date. It is important to estimate this parameter for application in PPH breeding, therefore, the aim of this study was to estimate the repeatability of overreach, term, and acuteness of PPHs. The study included records of 137 stallions and mares. All measurements were recorded in MP4 video format with a resolution of 1920 x 1080 megapixels and at 60 frames per second. All traits were measured three times (once per stride), and each trait was analyzed. KINOVEA software version 0.9.5 was used to analyze the measurements. A multivariate repeated measures animal model with sex effect was used to estimate the variance components for each trait using WOMBAT software. Repeatability was high in all traits (> 0.78; Table 1). The low standard error values close to 0.02 indicate high precision in all traits (0.904, 0.855, and 0.903 for overreach, term, and acuteness, respectively), being defined as the variance of the mean of “n” measurements as a percentage of the variance of one measurement, indicating that this parameter has little effect on the temporal environment of the three traits. These results demonstrate that previous training can influence the regularity of the gait, in agreement with the present study, in which only adult animals that have already had previous training were evaluated. Furthermore, our results corroborate that traits with high repeatability require few measurements (3 in this study) to obtain greater precision, and an increase in the number of measurements may be irrelevant for parameter estimation. In a breeding plan, this high precision may be due to an increase in the additive genetic variance, indicating that two or more measurements have greater heritability than a single measurement, allowing a better breeding value estimate to be obtained. Considering the high repeatability values found, this parameter can be used as an indicator of how effective the selection process can be, considering its relationship with heritability, as repeatability values are higher than heritability, due to the inclusion of permanent environmental variance (within-individual variance) in its estimation. High values also indicate that individuals tend to have consistent performance, and therefore, obtaining several measures for evaluation may be impractical. It can be concluded that overreach, term, and acuteness have high repeatability values, which allows these traits to respond better in a selection process and performance evaluation for subsequent competitions.

On-Field Rule Out of Mild Traumatic Brain Injury: Can Blood-Based Biomarker Testing Change Outcome of Major Sporting Events?

Mild traumatic brain injury (mTBI) is defined as a Glascow Coma Score of between 13 and 15. The diagnosis and rule out of individuals suffering from mTBI on an acute basis is imperfect and involves subjective measures. Serum biomarkers that exhibit narrow within-individual biological variation can be used for the early rule-out of mTBI, when baseline levels are compared during health. This is a descriptive study that applies published biological variation data of serum mTBI biomarkers for early rule out of sports-related injury. Laboratory tests such as glial fibrillary acidic protein, fatty acid binding protein 7, and phosphorylated protein enriched in astrocytes have low within-individual variances and are potential candidates. Aldolase C also rises early in blood but the biological variation is of this marker is currently unknown. The use of blood-based biomarkers, measured in real time using point-of-care testing devices when compared to a pre-competition baseline instead of a population-based reference interval, can provide early rule out of mTBI, and possibly enable on-field evaluations and a medical decision for a return to competition.

Individual flexibility in group foraging behaviour of reef manta rays (Mobula alfredi)

Flexibility in animal foraging strategies can increase overall feeding efficiency for individuals. For example, group foraging can increase the efficiency of resource exploitation; conversely solo foraging can reduce intraspecific competition, particularly at low resource densities. The cost–benefit trade-off of such flexibility is likely to differ within and among individuals. Reef manta rays (Mobula alfredi) are large filter-feeding elasmobranchs that often aggregate to feed on ephemeral upwellings of zooplankton. Over three years in the Maldives, we free-dived to film 3106 foraging events involving 343 individually identifiable M. alfredi. Individuals fed either solo or in groups with a clear leader plus between one and eight followers. M. alfredi were significantly more likely to forage in groups than solo at high just prior to high tide and when aggregations were larger. Within aggregations, individuals foraged in larger groups when more food was available, and when the overall aggregations were relatively large suggesting that foraging in large groups was more beneficial when food is abundant, and the costs of intraspecific competition were outweighed by the efficiency resulting from group foraging strategies. Females, the larger sex, were more likely to lead foraging groups than males. The high within-individual variance (over 70%), suggested individuals were unpredictable across all foraging behaviours, thus individual M. alfredi cannot be classified into foraging types or specialists. Instead, each individual was capable of considerable behavioural flexibility, as predicted for a species reliant on spatially and temporally ephemeral resources.Significance statementReef manta rays (Mobula alfredi), listed as “Vulnerable” on the IUCN Red List, are at risk from targeted and by-catch fisheries due to their slow life history and aggregative behaviour. M. alfredi feed together in aggregations on short-lived glut of microscopic zooplankton. Over three years in the Maldives, we filmed 3106 foraging events involving 343 individually identifiable M. alfredi. Manta rays were more likely to forage in groups than solo just prior to high tide and when aggregations were larger, attracted by the influx of zooplankton. Foraging groups included more individuals when plankton was more abundant. However, individuals flipped between solo and group foraging and did not specialise. Foraging groups were most often led by females, the larger sex. Individuals were very flexible in how they foraged, which makes sense for a species that relies on a food source that varies enormously in when, where and for how long it is available. Understanding manta ray foraging behaviour will help conservation management efforts and predict their responses to climate change.

Are there sex differences in the variability of fasting metabolism?

There is evidence across species and across many traits that males display greater between-individual variance. In contrast, (premenopausal) females display large within-individual variance in sex hormone concentrations, which can increase within-individual variance in many other parameters. The latter may contribute to the lower representation of females in metabolic research. This study is a pooled secondary analysis of data from seven crossover studies to investigate the between-individual and the within-individual variance in fasting plasma metabolites, resting metabolic rate (RMR), and body mass. Females demonstrated higher within-individual variability of plasma 17β-estradiol [coefficient of variation (CV): 15 ± 15% for males vs. 38 ± 34% for females, P < 0.001] and progesterone concentrations (CV: 13 ± 11% for males vs. 52 ± 51% for females, P < 0.001) but there were no meaningful differences in the variability of plasma glucose (CV: 4 ± 3% for males vs. 5 ± 5% for females), insulin, lactate, triglycerides (CV: 15 ± 9% for males vs. 15 ± 10% for females), and esterified fatty acid concentrations or in RMR and body mass (CV: 0.43 ± 0.34% for males vs. for 0.42 ± 0.33% females; P > 0.05 for all outcomes). Males displayed higher between-individual variance in RMR compared with females (SD: 224 kcal·day-1 for males vs. 151 kcal·day-1 for females). In conclusion, these data do not provide evidence that females show greater within-individual variability in many fasting metabolic variables, RMR, or body mass compared with males. We conclude that including females in metabolic research is unlikely to introduce greater within-individual variance when using the recruitment and control procedures described in these studies.NEW & NOTEWORTHY To investigate the within-individual variability in metabolic parameters in males and females, we performed a pooled secondary analysis of fasting blood samples, resting metabolic rate, and body mass from seven crossover studies. We found a greater day-to-day variation in 17β-estradiol and progesterone in females compared with males but no meaningful difference in within-individual variability of fasting plasma glucose, insulin, lactate, triglycerides, NEFA, resting metabolic rate, or body mass between females and males.

Sexual Selection Increases Male Behavioral Consistency in Drosophila melanogaster.

AbstractSexual selection has been suggested to influence the expression of male behavioral consistency. However, despite predictions, direct experimental support for this hypothesis has been lacking. Here, we investigated whether sexual selection altered male behavioral consistency in Drosophila melanogaster-a species with both pre- and postcopulatory sexual selection. We took 1,144 measures of locomotor activity (a fitness-related trait in D. melanogaster) from 286 flies derived from replicated populations that have experimentally evolved under either high or low levels of sexual selection for >320 generations. We found that high sexual selection males were more consistent (decreased within-individual variance) in their locomotor activity than male conspecifics from low sexual selection populations. There were no differences in behavioral consistency between females from the high and low sexual selection populations. Furthermore, while females were more behaviorally consistent than males in the low sexual selection populations, there were no sex differences in behavioral consistency in high sexual selection populations. Our results demonstrate that behavioral plasticity is reduced in males from populations exposed to high levels of sexual selection. Disentangling whether these effects represent an evolved response to changes in the intensity of selection or are manifested through nongenetic parental effects represents a challenge for future research.

The effect of repeated measurements and within-individual variance on the estimation of heritability: a simulation study

The estimation of heritability is a common practice in the field of ecology and evolution. Heritability of the traits is often estimated using one single measurement per individual, although many traits (especially behavioural and physiological traits) are characterized by large within-individual variance, and ideally a large number of within individual measurements can be obtained. Importantly, the effect of the within-individual variance and the rate at which this variance is sampled on the estimation of heritability has not been thoroughly tested. We fill this gap of knowledge with a simulation study, and assess the effect of within- and between-individual sample size, and the true value of the variance components on the estimation of heritability. In line with previous studies we found that the accuracy and precision of heritability estimation increased with sample size and accuracy with higher values of additive genetic variance. When the sample size was above 500 accuracy and power of heritability estimates increased in the models including repeated measurements, especially when within-individual variance was high. We thus suggest to use a sample of more than 100 individuals and to include more than two repeated measurements per individual in the models to improve estimation when investigating heritability of labile traits.Significance statementHeritability reflects the part of the trait’s phenotypic variation underlined by genetic variation. Despite the difficulties of heritability calculation (high number of individuals is needed with known relatedness), it is a widely used measure in evolutionary studies. However, not every factor potentially affecting the quality of heritability estimation is well understood. We thus investigated with a comprehensive simulation study how the number of repeated measurements per individuals and the amount of within-individual variation influence the goodness of heritability estimation. We found that although the previously described effect of the number of studied individuals was the most important, including repeated measurements also improved the reliability of the heritability estimates, especially when within-individual variation was high. Our results thus highlight the importance of including repeated measurements when investigating the heritability of highly plastic traits, such as behavioural or physiological traits.

Among- and Within-Individual Variance in Metabolic Thermal Reaction Norms.

AbstractIn ectotherms, temperature has a strong effect on metabolic rate (MR), yet the extent to which the thermal sensitivity of MR varies among versus within individuals is largely unknown. This is of interest because significant among-individual variation is a prerequisite for the evolution of metabolic thermal sensitivity. Here, we estimated the repeatability (R) of the thermal sensitivity of MR in individual virgin, adult male Drosophila melanogaster () by taking repeated overnight measures of their MRs at two temperatures (~24°C and ~27°C). At the population level, thermal sensitivity decreased with locomotor activity, and older individuals showed a higher thermal sensitivity of MR than younger individuals. Taking these effects (and body mass) into account, we detected significant repeatability in both the centered intercept () and the slope () of the metabolic thermal reaction norms, which respectively represent average MR and thermal sensitivity of MR. Furthermore, individuals with a higher overall MR also displayed greater increases in MR as temperature increased from ~24°C to ~27°C (). Average MR and thermal sensitivity of MR were also positively correlated within individuals (). Our study represents a point of departure for future larger studies, in which more complex protocols (e.g., wider temperature range, breeding design) can be applied to quantify the causal components of variation in thermal sensitivity that are needed to make accurate predictions of adaptive responses to global warming.

Weak correlations between visual abilities in healthy older adults, despite long-term performance stability

Using batteries of visual tests, most studies have found that there are only weak correlations between the performance levels of the tests. Factor analysis has confirmed these results. This means that a participant excelling in one test may rank low in another test. Hence, there is very little evidence for a common factor in vision. In visual aging research, cross-sectional studies have repeatedly found that healthy older adults’ performance is strongly deteriorated in most visual tests compared to young adults. However, also within the healthy older population, there is no evidence for a visual common factor. To investigate whether the weak between-tests correlations are due to fluctuations in individual performance throughout time, we conducted a longitudinal study. Healthy older adults performed a battery of eight visual tests, with two re-tests after approximately four and seven years. Pearson’s, Spearman’s and intraclass correlations of most visual tests were significant across the three testing, indicating that the tests are reliable and individual differences are stable across years. Yet, we found low between-tests correlations at each visit, which is consistent with previous studies finding no evidence for a visual common factor. Our results exclude the possibility that the weak correlations between tests are due to high within-individual variance across time.

Selection for environmental variance shifted the gut microbiome composition driving animal resilience

BackgroundUnderstanding how the host’s microbiome shapes phenotypes and participates in the host response to selection is fundamental for evolutionists and animal and plant breeders. Currently, selection for resilience is considered a critical step in improving the sustainability of livestock systems. Environmental variance (VE), the within-individual variance of a trait, has been successfully used as a proxy for animal resilience. Selection for reduced VE could effectively shift gut microbiome composition; reshape the inflammatory response, triglyceride, and cholesterol levels; and drive animal resilience. This study aimed to determine the gut microbiome composition underlying the VE of litter size (LS), for which we performed a metagenomic analysis in two rabbit populations divergently selected for low (n = 36) and high (n = 34) VE of LS. Partial least square-discriminant analysis and alpha- and beta-diversity were computed to determine the differences in gut microbiome composition among the rabbit populations.ResultsWe identified 116 KEGG IDs, 164 COG IDs, and 32 species with differences in abundance between the two rabbit populations studied. These variables achieved a classification performance of the VE rabbit populations of over than 80%. Compared to the high VE population, the low VE (resilient) population was characterized by an underrepresentation of Megasphaera sp., Acetatifactor muris, Bacteroidetes rodentium, Ruminococcus bromii, Bacteroidetes togonis, and Eggerthella sp. and greater abundances of Alistipes shahii, Alistipes putredinis, Odoribacter splanchnicus, Limosilactobacillus fermentum, and Sutterella, among others. Differences in abundance were also found in pathways related to biofilm formation, quorum sensing, glutamate, and amino acid aromatic metabolism. All these results suggest differences in gut immunity modulation, closely related to resilience.ConclusionsThis is the first study to show that selection for VE of LS can shift the gut microbiome composition. The results revealed differences in microbiome composition related to gut immunity modulation, which could contribute to the differences in resilience among rabbit populations. The selection-driven shifts in gut microbiome composition should make a substantial contribution to the remarkable genetic response observed in the VE rabbit populations.6gPh8oS1o7RBSMdC-HYk4TVideo

The Leydig cell biomarker INSL3 as a predictor of age-related morbidity: Findings from the EMAS cohort.

Insulin-like peptide 3 (INSL3) is a constitutive hormone secreted in men by the mature Leydig cells of the testes. It is an accurate biomarker for Leydig cell functional capacity, reflecting their total cell number and differentiation status. To determine the ability of INSL3 to predict hypogonadism and age-related morbidity using the EMAS cohort of older community-dwelling men. Circulating INSL3 was assessed in the EMAS cohort and its cross-sectional and longitudinal relationships to hypogonadism, here defined by testosterone (T) <10.5nmol/l, and a range of age-related morbidities determined by correlation and regression analysis. While INSL3 is an accurate measure of primary hypogonadism, secondary and compensated hypogonadism also indicate reduced levels of INSL3, implying that testicular hypogonadism does not improve even when LH levels are increased, and that ageing-related hypogonadism may combine both primary and secondary features. Unadjusted, serum INSL3, like calculated free testosterone (cFT), LH, or the T/LH ratio reflects hypogonadal status and is associated with reduced sexual function, bone mineral density, and physical activity, as well as increased occurrence of hypertension, cardiovascular disease, cancer, and diabetes. Using multiple regression analysis to adjust for a range of hormonal, anthropometric, and lifestyle factors, this relationship is lost for all morbidities, except for reduced bone mineral density, implying that INSL3 and/or its specific receptor, RXFP2, may be causally involved in promoting healthy bone metabolism. Elevated INSL3 also associates with hypertension and cardiovascular disease. When unadjusted, INSL3 in phase 1 of the EMAS study was assessed for its association with morbidity in phase 2 (mean 4.3 years later); INSL3 significantly predicts 7 out of 9 morbidity categories, behaving as well as cFT in this regard. In contrast, total T was predictive in only 3 of the 9 categories. Together with its low within-individual variance, these findings suggest that assessing INSL3 in men could offer important insight into the later development of disease in the elderly.

Estimating heritability of song considering within-individual variance in a wild songbird: The collared flycatcher

Heritable genetic variation is a prerequisite for adaptive evolution; however, our knowledge about the heritability of plastic traits, such as behaviors, is scarce, especially in wild populations. In this study, we investigated the heritability of song traits in the collared flycatcher (Ficedula albicollis), a small oscine passerine with complex songs involved in sexual selection. We recorded the songs of 81 males in a natural population and obtained various measures describing the frequency, temporal organization, and complexity of each song. As we had multiple songs from each individual, we were able to statistically account for the first time for the effect of within-individual variance on the heritability of song. Heritability was calculated from the variance estimates of animal models relying on a genetic similarity matrix based on Single Nucleotide Polymorphism screening. Overall, we found small additive genetic variance and heritability values in all song traits, highlighting the role of environmental factors in shaping bird song.

Living on the edge: morphological and behavioral adaptations to a marginal high-elevation habitat in an arboreal mammal.

Habitats are characterized by different local environmental conditions that influence both behavior and morphology of species, which can result in habitat-dependent phenotypic differences among animals living in heterogeneous environments. We studied 3 alpine populations of Eurasian red squirrels (Sciurus vulgaris), 1 living in a marginal high-elevation habitat at the edge of the species' altitudinal distribution, and 2 occurring in higher-quality habitats. Here, we investigated whether squirrels living in the marginal area differed in 2 morphological parameters (body size and body mass) and/or in the expression of 4 personality traits estimated with an open field test and a mirror image stimulation test (activity, exploration, activity-exploration, and social tendency). Furthermore, we tested whether within-individual variance of the traits (behavioral plasticity) was higher in the edge habitat. Male squirrels in the edge habitat were smaller and weighed less than in the other study areas, while among females, size-habitat relationships were less marked. These sex-specific patterns were explained by a strong association between body mass and reproductive success in female squirrels. Squirrels in the marginal habitat were more active, explorative, and had a more social personality than in the other habitats. However, in contrast to our predictions, behavioral plasticity was smaller in the marginal habitat, but only for the trait exploration. Our results suggest that animals choose the habitat that best fits their personality, and that habitat-related differences in selective pressures may shape animals' morphology.

A paradigm shift in learning strategy research: Illustration and example of a within-person examination.

Although research on learning strategies has contributed to improving learners' performance, most studies only described the characteristics of learners. These approaches have focused on inter-individual differences and their correlations, making it difficult to examine the process of selecting learning strategies in individuals. To examine the correlations within individual learners, such as the variables that influence the use of a strategy by a specific learner, two surveys were conducted in this study. Perceived cost and perceived benefit of using each strategy as the influential variables on choosing a strategy were adopted with reference to decision-making research. Perceived cost consisted of the time perspective for learning outcomes by the learning strategy and the method of learning strategy implementation. In Study 1, cognitive strategies that have a direct impact on the acquisition of learning content were measured, and metacognitive strategies that monitor and regulate one's own learning behavior were examined in Study 2. The variables were treated for each individual to calculate intra-class and within-person correlations, although data were collected using a typical questionnaire survey in learning strategy studies. Hierarchical Bayesian modeling was conducted with use of the learning strategy as the objective variable and the perceived costs and benefits as explanatory variables. The analysis revealed that, in terms of cognitive strategies (Study 1), learners avoided using learning strategies perceived to have a high cost in the short term; however, they used cognitive strategies regardless of the circumstances. Furthermore, regarding metacognitive strategies (Study 2), students avoided using learning strategies that they perceived as costly to use in the short term and used them only when they suited the circumstances. On the other hand, in metacognitive strategies (Study 2), students avoided using learning strategies that they perceived as costly to use for the next test and use as appropriate for the situation. Focusing on within-individual variance and correlation made the interpretation of factors that influence the choice of learning strategies more intuitive and provided more suggestions for educational practice.

Influence on the adult male Leydig cell biomarker insulin-like peptide 3 of maternal exposure to estrogenic and anti-androgenic endocrine disrupting compound mixtures: A retrospective study.

Insulin-like peptide 3 (INSL3) is a peptide biomarker secreted specifically by the mature Leydig cells of the testes. It is constitutive, has low within-individual variance, and effectively measures the functional capacity of Leydig cells to make testosterone. In young adult men there is a large 10-fold range of serum INSL3 concentration, persisting into old age, and implying that later hypogonadal status might be programmed in early life. To determine whether maternal exposure to environmental endocrine disrupting compounds (EDCs) influences adult serum INSL3 concentration, using a retrospective paradigm, INSL3 was measured in young adult male rats (80-90 days) from the F1 generation of females maternally exposed to varied doses of bisphenol A (BPA), butylparaben, epoxiconazole, and fludioxonil as single compounds, as well as estrogenic and anti-androgenic mixtures of BPA and butylparaben, and di(2-ethylhexyl) phthalate and procymidone respectively. A mixture of BPA and butylparaben significantly reduced circulating INSL3 concentration in adult male progeny. The remaining compounds or mixtures tested, though sufficient to induce other effects in the F1 generation were without significant effect. Maternal exposure to low concentrations of some EDCs may be a contributing factor to the variation in the Leydig cell biomarker INSL3 in young adulthood, though caution is warranted translating results from rats to humans.

Endocrine flexibility.

Endocrine flexibility.

Sources of Variability in Serum Lipidomic Measurements and Implications for Epidemiologic Studies.

Epidemiological studies using lipidomic approaches can identify lipids associated with exposures and diseases. We evaluated the sources of variability of lipidomic profiles measured in blood samples and the implications when designing epidemiologic studies. We measured 918 lipid species in nonfasting baseline serum from 693 participants in the Prostate, Lung, Colorectal, and Ovarian Cancer Screening Trial, with 570 participants having serial blood samples separated by 1-5 years and 72 blinded replicate quality control samples. Blood samples were collected during 1993-2006. For each lipid species, we calculated the between-individual, within-individual, and technical variances, and we estimated the statistical power to detect associations in case-control studies. The technical variability was moderate, with a median intraclass correlation coefficient of 0.79. The combination of technical and within-individual variances accounted for most of the variability in 74% of the lipid species. For an average true relative risk of 3 (comparing upper and lower quartiles) after correction for multiple comparisons at the Bonferroni significance threshold (α= 0.05/918= 5.45×10-5), we estimated that a study with 500, 1,000, and 5,000 total participants (1:1 case-control ratio) would have 19%, 57%, and 99% power, respectively. Epidemiologic studies examining associations between lipidomic profiles and disease require large samples sizes to detect moderate effect sizes associations.

A quantitative genetics perspective on the body-mass scaling of metabolic rate

Widely observed allometric scaling (log-log slope<1) of metabolic rate (MR) with body mass (BM) in animals has been frequently explained using functional mechanisms, but rarely studied from the perspective of multivariate quantitative genetics. This is unfortunate, given that the additive genetic slope (bA) of the MR-BM relationship represents the orientation of the 'line of least genetic resistance' along which MR and BM may most likely evolve. Here, we calculated bA in eight species. Although most bA values were within the range of metabolic scaling exponents reported in the literature, uncertainty of each bA estimate was large (only one bA was significantly lower than 3/4 and none were significantly different from 2/3). Overall, the weighted average for bA (0.667±0.098 95% CI) is consistent with the frequent observation that metabolic scaling exponents are negatively allometric in animals (b<1). Although bA was significantly positively correlated with the phenotypic scaling exponent (bP) across the sampled species, bP was usually lower than bA, as reflected in a (non-significantly) lower weighted average for bP (0.596±0.100). This apparent discrepancy between bA and bP resulted from relatively shallow MR-BM scaling of the residuals [weighted average residual scaling exponent (be)=0.503±0.128], suggesting regression dilution (owing to measurement error and within-individual variance) causing a downward bias in bP. Our study shows how the quantification of the genetic scaling exponent informs us about potential constraints on the correlated evolution of MR and BM, and by doing so has the potential to bridge the gap between micro- and macro-evolutionary studies of scaling allometry.

Repeatability of endocrine traits and dominance rank in female guinea pigs

BackgroundGlucocorticoids (e.g. cortisol) are associated with variation in social behavior, and previous studies have linked baseline as well as challenge-induced glucocorticoid concentrations to dominance status. It is known that cortisol response to an acute challenge is repeatable and correlates to social behavior in males of many mammal species. However, it is unclear whether these patterns are also consistent for females. The aim of this study was to investigate whether baseline and response cortisol concentrations are repeatable in female guinea pigs (Cavia aperea f. porcellus) and whether dominance rank is stable and correlated to baseline cortisol concentration and/or cortisol responsiveness.ResultsOur results show that cortisol responsiveness (after 1 h: R = 0.635, 95% CI = 0.229, 0.927; after 2 h: R = 0.764, 95% CI = 0.433, 0.951) and dominance rank (R = 0.709, 95% CI = 0.316, 0.935) of females were significantly repeatable after six weeks but not correlated. Baseline cortisol was not repeatable (R = 0, 95% CI = 0, 0.690) and also did not correlate to dominance rank. Furthermore, the difference in repeatability estimates of baseline and response values was due to high within-individual variance of baseline cortisol concentration; the amount of between-individual variance was similar for baseline cortisol and the two measures of cortisol responsiveness.ConclusionsFemales occupying different dominance ranks did not have long-term differences in cortisol concentrations, and cortisol responsiveness does not seem to be significantly involved in the maintenance of dominance rank. Overall, this study reveals the remarkable stability of cortisol responsiveness and dominance rank in a female rodent, and it remains an open question whether the magnitude of cortisol responsiveness is adaptive in social contexts for females.

Comparison of isotope ratio mass spectrometry and cavity ring-down spectroscopy procedures and precision of the doubly labeled water method in different physiological specimens.

This study determines if saliva collection procedures for the doubly labeled water (DLW) method, used for measuring total energy expenditure (TEE), are comparable to urine and plasma collection. Both the cavity ring-down spectroscopy (CRDS) and isotope ratio mass spectrometry (IRMS) analyses techniques are compared. Saliva specimens were collected from participants for the DLW method. The specimens were collected under different conditions: after consumption of tap water, after chewing gum, and during exposure to conditions of high and low relative humidity. The isotopes in saliva were compared with simultaneous plasma and urine collection. TEE calculated from saliva and analyzed using CRDS was compared to that of plasma analyzed using IRMS. The within-individual variances were not significantly different between the saliva specimens (0.4‰) and plasma (0.3‰). After the oral dose of DLW, the saliva specimens displayed a shorter equilibration time to urine. When participants consumed 500 mL of tap water, the enrichment of saliva specimens reached a new plateau value faster than urine. Saliva collection exposed to high ambient humidity conditions was slightly less enriched as compared to low-humidity conditions while urine enrichment was unaffected. In contrast, whereas the within-individual effects of gum chewing during saliva collection on 18 O were unaffected, the abundance of 2 H in saliva was slightly lower after chewing the gum. The within-individual difference between TEE calculated from saliva and that calculated from plasma analyzed using IRMS did not differ from zero, and the standard deviation was not different from that predicted by a propagation of error analysis based on analytical error alone. Our findings support using saliva specimens for the DLW method. The analysis of plasma and urine, however, requires reducing the memory effect resulting from contaminants. Also, it should be performed in a manner that minimizes exposure to air where specimens may be exposed to evaporation or contamination from water vapor during sampling.

Test-Retest Reliability and Consistency of HVPG and Impact on Trial Design: A Study in 289 Patients from 20 Randomized Controlled Trials.

Portal hypertension (PH) is a major driver for cirrhosis complications. Portal pressure is estimated in practice by the HVPG. The assessment of HVPG changes has been used for drug development in PH. This study aimed at quantifying the test-retest reliability and consistency of HVPG in the specific context of randomized controlled trials (RCTs) for the treatment of PH in cirrhosis and its impact on power calculations for trial design. We conducted a search of published RCTs in patients with cirrhosis reporting individual patient-level data of HVPG at baseline and after an intervention, which included a placebo or an untreated control arm. Baseline and follow-up HVPGs in the control groups were extracted after digitizing the plots. We assessed reliability and consistency and the potential impact of study characteristics. We retrieved a total of 289 before and after HVPG measurements in the placebo/untreated groups from 20 RCTs. The time span between the two HVPG measurements ranged between 20minutes and 730days. Pre-/post-HVPG variability was lower in studies including only compensated patients; therefore, modeled sample size calculations for trials in compensated cirrhosis were lower than for decompensated cirrhosis. A higher proportion of alcohol-associated cirrhosis and unicentric trials was associated with lower differences between baseline and follow-up measurements. The smallest detectable difference in an individual was 26% and 30% in compensated and decompensated patients, respectively. The test-retest reliability of HVPG is overall excellent. Within-individual variance was higher in studies including higher proportions of decompensated patients. These findings should be taken into account when performing power analysis for trials based on the effects on HVPG or when considering HVPG as a tool to guide therapy of PH.