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Stier, A.

Publications and source records attributed to Stier, A..

At least 19 recordsLinked to original sources

The king of stress? Exploring the physiological resilience and resistance of adult king penguins to chronic glucocorticoid exposure

To better understand how animals cope with increasingly variable and challenging environments, there is a need to study how prolonged exposure to elevated glucocorticoid hormones (i.e. one mediator of the stress response) affects their physiology. While glucocorticoid elevation is known to increase oxidative stress and accelerate cellular ageing, there is evidence that king penguins (Aptenodytes patagonicus) can prevent oxidative stress during acute stress exposure, suggesting that species may differ in their sensitivity to glucocorticoids downstream negative effects. As king penguins thrive in a seemingly harsh environment, we hypothesized that they may be able to limit the deleterious effects usually associated with chronic glucocorticoid elevation, either through resistance (i.e. prevention of downstream negative effect) or resilience (i.e. rapid recovery following transient negative effect). To test this hypothesis, we experimentally elevated corticosterone levels in incubating king penguins and quantified treatment effects on a suite of physiological traits at multiple time points across incubation and early chick-rearing, up to ca. 2 months after implantation. Corticosterone-treated individuals showed a prolonged increase in corticosterone and decrease in body condition, confirming our treatment likely mimicked sustained stress exposure. Heterophil-to-lymphocyte ratio was only increased transiently, and there was no clear evidence that treatment influenced oxidative stress or telomere length maintenance. Plasma energy metabolites were mainly affected early after implantation, with rapid recovery over time. Overall, our results suggest that adult king penguins show at least moderate resistance and resilience to chronic corticosterone elevation, especially in preventing cellular integrity loss, though at-sea physiological effects remain to be determined.

physiology↗

Cortical Thickness and Curvature in Autism and ADHD: A Mega-Analysis

BackgroundExisting evidence suggests cortical morphometric alterations occur in people with autism and ADHD. However, these findings remain tentative due to small sample sizes, heterogeneous imaging pipelines, varied statistical approaches, and limited harmonization across acquisition sites. Few studies have applied standardized processing to large, clinically enriched datasets or addressed site-related batch effects. MethodsWe leveraged six large-scale brain imaging datasets (n = 9,647; male=5,835; female=3,812; ages 5-64 years), including 1,533 individuals with ADHD, 1,080 with autism spectrum disorder, and 7,034 matched controls. All imaging data were processed using the validated ABCD-HCP pipeline, with cortical parcellation into 360 regions based on the Human Connectome Project (HCP) atlas, and ComBat harmonization was applied to account for variability across 67 acquisition sites. Group-level differences in cortical thickness and sulcal curvature were examined with ANCOVAs, controlling for covariates and using Bonferroni correction for multiple comparisons. ResultsOur analyses revealed distinct neuroanatomical signatures for both autism and ADHD. Individuals with autism exhibited regionally thinner cortex and curvature alterations particularly in the Cingulo-Opercular network. In contrast, individuals with ADHD displayed regionally thicker cortex, particularly in the default mode and somatomotor networks, alongside curvature differences. Control participants showed intermediate patterns, suggesting that autism and ADHD may represent diverging extremes of cortical maturation. ConclusionsCortical thickness and curvature emerge as potential biomarkers that can advance understanding of neurodevelopmental conditions and disentangle heterogeneity across diagnostic groups. These findings highlight the value of harmonized, large-scale, standardized analyses for resolving inconsistencies in the literature.

neuroscience↗

Vaccination against H5 HP influenza virus leads to persistent immune response in wild king penguins

Since 2021, the panzootic of high pathogenicity avian influenza (HPAI) represents an increasing threat to wild vertebrate populations. In this context, recent vaccines developed for poultry could provide tools for the conservation of wild endangered birds populations. The king penguin (Aptenodytes patagonicus), a long-lived seabird breeding in dense colonies with an extended chick-rearing period, was identified as a possible surrogate species for a vaccination trial in a sub-Antarctic natural setting. We investigate here the immune response of king penguin chicks to a self-amplifying mRNA vaccine against a H5 HPAI clade 2.3.4.4b protein. Thirty chicks were vaccinated (primo- and boost-injections), 20 were kept as controls. Along 250 days of monitoring, vaccinated chicks showed a high and persistent immune response, granting a strong seroneutralisation capacity against the virus, up to fledging. No adverse effects were observed. The screening for antibodies against unspecific avian influenza viruses suggested that no natural infection occurred over the entire trial. The emergence of HPAI in the Southern Indian Ocean in October 2024 highlights the timeliness of such experimental tests. Our results show the vaccine could provide a powerful tool for mitigation and emphasises the need for studies considering ethical and practical issues of vaccination for wildlife conservation.

ecology↗

Cannot be male without complex I? Alteration in cellular metabolism involved in cytoplasmic male sterility in snail Physa acuta

Cytoplasmic male sterility (CMS) is a well-known example of mitonuclear genetic conflict over sex determination in hermaphrodite plants, where mitochondrial genes maternally inherited sterilize the male function while biparental inherited nuclear genes restore it. CMS has been recently discovered in wild animals, in the freshwater snail Physa acuta. In this species, CMS is associated with two extremely divergent mitogenomes D and K compared to the classical mitogenome N. D individuals are male-steriles, while male fertility is restored in K individuals. We hypothesized that the extreme divergence of mitogenomes associated with CMS can impact mitochondrial aerobic metabolism, a necessary process in eukaryotic organisms by which energy is transduced from food to ATP. Our results suggest that CMS might be associated with an alteration of co-encoded complex I in D male-sterile individuals, although partly compensated by the nuclear-encoded complex II. K restored hermaphrodites have an unaffected complex I respiration, but exhibit higher mitochondrial proton leak and higher relative anaerobic contribution to cellular metabolism, suggesting an energy cost of bearing CMS and restorer genes, which could underlie the reduced growth of K mitotype. How complex I alteration might induce male-sterility remains to be determined, but could be linked to oxidative stress or a defect in ATP-synthesis rate.

evolutionary biology↗

Warming up to a new coat: moulting king penguins exhibit hyperthermia and increased peripheral heat loss

Penguins are among the most specialized thermoregulators on the planet, however, the same adaptations that maximize heat retention underwater likely hinder heat dissipation on land, possibly creating dangerous thermoregulatory challenges when encountering warming terrestrial habitats. Penguins are subject to strictly terrestrial phases, such as moulting, when metabolic heat production, insulation, and energetic constraints are heightened. We assessed thermoregulation in moulting captive king penguins (Aptenodytes patagonicus) using simultaneous measurements of core and surface temperatures to test two hypotheses. Under the thermal challenge hypothesis, an initial rise in heat dissipation effort (i.e., increased peripheral vasomotion) followed by a rise in core temperature would indicate failure to prevent hyperthermia. Under the warm-up hypothesis, an initial rise of core temperature concomitant or followed by an increase of peripheral vasomotion would indicate regulated hyperthermia, possibly to accelerate feather development. Core and surface temperatures increased drastically but concomitantly during moult, providing tentative support for the warm-up hypothesis. Moulting penguins did not pant, suggesting that peripheral heat dissipation was sufficient to regulate moulting-induced hyperthermia. Core and subcutaneous temperatures in wild individuals resembled patterns measured in captivity, despite lower heat load and additional options for behavioural thermoregulation. These results indicate that hyperthermia is prevalent in moulting king penguins, and documenting the timing of temperature changes provides novel insights for the moulting physiology of penguins. Because moulting-induced hyperthermia may contribute to heat load, we caution that moulting may increase the susceptibility of wild penguins to heat stress, especially as regions near the poles warm at a disproportionately rapid rate.

physiology↗

The effects of early-life ambient temperature on the gut microbiome of a wild bird? an experimental approach

Vertebrate gut microbiome has significant effects on host development, health, and fitness. Multiple external factors contribute to gut microbiome variation, and the role of ambient temperature has gained increasing attention. Yet, temperature effects are often tested in extremes and in captive systems. We experimentally studied the effect of subtle temperature decreases during post-natal development on gut microbiome diversity and composition in wild pied flycatchers (Ficedula hypoleuca). We also performed partial cross-fostering to study the relative contribution of genetic and rearing environment on microbiome. Nest-box cold treatment did not influence gut microbiome diversity or composition, which may be due to the small temperature change, ontogenetic stage, or other factors, such as diet, causing large variation in the data. Rearing environment explained more of the variation in gut microbiome than genetic background, but the variance explained was relatively small. Future studies need to further address the drivers of the large intraspecific variation in microbiome in natural populations.

ecology↗

Mind the polar sun: Solar radiations trigger frequent heat stress in breeding king penguins, despite relatively cool air temperatures.

Polar and sub-polar animals evolved to thrive in cold climates and may thus be particularly sensitive to rising temperatures associated with climate change. Penguins may be especially vulnerable, due to their dual habitat, alternating between foraging in cold waters and breeding/moulting on an increasingly warm land. Here, we characterized heat stress occurrence in breeding king penguins through behavioural observations (e.g. panting occurrence) and body temperature measurements. We observed that behavioural signs of heat stress are frequent in king penguins breeding in the sub-Antarctic region (> 20% of observations at mid-day), and that subcutaneous temperatures increase under high heat load, especially in penguins observed panting. Subcutaneous and core body temperatures were moderately correlated and both increased with heat load. Yet, their responses were not parallel since core body temperature is markedly less sensitive to heat load than subcutaneous temperature. Air temperature alone was a poor predictor of heat stress occurrence, whereas the combination of high solar radiation, low wind speed and high air temperatures provided the strongest predictive power. Finally, reproductive failures were more likely to occur on warmer days, suggesting that heat stress may have significant sublethal effects on adults that could ultimately affect population dynamics. O_FIG O_LINKSMALLFIG WIDTH=150 HEIGHT=200 SRC="FIGDIR/small/611977v3_ufig1.gif" ALT="Figure 1"> View larger version (95K): org.highwire.dtl.DTLVardef@26db91org.highwire.dtl.DTLVardef@119461dorg.highwire.dtl.DTLVardef@4a1d8forg.highwire.dtl.DTLVardef@1da9d96_HPS_FORMAT_FIGEXP M_FIG C_FIG

evolutionary biology↗

Changes in surface temperatures reveal the thermal challenge associated with catastrophic moult in captive Gentoo penguins

Once a year, penguins undergo a catastrophic moult replacing their entire plumage during a fasting period on land or on sea-ice during which time individuals can lose 45% of their body mass. In penguins, new feather synthesis precedes the loss of old feathers leading to an accumulation of two feathers layers (double coat) before the old plumage is shed. We hypothesize that the combination of the high metabolism required for new feathers synthesis and the potentially high thermal insulation linked to the double coat could lead to a thermal challenge requiring additional peripheral circulation to thermal windows to dissipate extra-heat. To test this hypothesis, we measured the surface temperature of different body regions of captive Gentoo penguins (Pygoscelis papua) throughout the moult under constant environmental conditions. The surface temperature of the main body trunk decreased during the initial stages of the moult, therefore suggesting a higher thermal insulation. On the opposite, the periorbital region, a potential proxy of core temperature in birds, increased during these same early moulting stages. The surface temperature of bill, flipper and foot (thermal windows) tended to initially increase during the moult period, highlighting the likely need for extra heat dissipation in moulting penguins. These results raise questions regarding the thermoregulatory capacities of wild penguins during the challenging period of moulting on land in the current context of global warming.

physiology↗

Mitonuclear interactions impact aerobic metabolism in hybrids and may explain mitonuclear discordance in young, naturally hybridizing bird lineages

Understanding genetic incompatibilities and genetic introgression between incipient species are major goals in evolutionary biology. Mitochondrial genes evolve rapidly and exist in dense gene networks with coevolved nuclear genes, suggesting that mitochondrial respiration may be particularly susceptible to disruption in hybrid organisms. Mitonuclear interactions have been demonstrated to contribute to hybrid disfunction between deeply divergent taxa crossed in the laboratory, but there are few empirical examples of mitonuclear interactions between younger lineages that naturally hybridise. Here we use experimental crosses and high resolution respirometry to provide the first evidence in a bird that inter-lineage mitonuclear interactions impact mitochondrial aerobic metabolism. Specifically, respiration capacity of the two paternal backcrosses (with mismatched mito-nuclear combinations) differ from one another, although they do not differ to the parental groups or maternal backcrosses as we would expect of mitonuclear disruptions. In the wild hybrid zone between these subspecies the mitochondrial cline centre is shifted west of the nuclear cline centre, which is consistent with the direction of our experimental results. Our results therefore demonstrate asymmetric mitonuclear interactions that impact the capacity of cellular mitochondrial respiration and may help to explain the geographic discordance between mitochondrial and nuclear genomes observed in the wild.

evolutionary biology↗

Early-life environmental effects on mitochondrial aerobic metabolism: an experimental brood size manipulation in wild great tits

Parental care (including postnatal provisioning) is a major component of the offsprings early-life environment. In avian species, the number of chicks in the nest and subsequent sibling competition for food are known to affect chicks growth, leading in some cases to long-lasting effects for the offspring. Because of its central role in converting energy, variation in the offsprings mitochondrial metabolism could be an important pathway underlying variation in growth patterns. Here, we performed a brood size manipulation in great tits (Parus major) to unravel its impact on offsprings mitochondrial metabolism and reactive oxygen species (ROS) production in red blood cells. We investigated the effects of brood size on chicks growth and survival, and tested for long-lasting effects on juvenile mitochondrial metabolism and phenotype. As expected, chicks raised in reduced broods had a higher body mass compared to enlarged and control groups. However, mitochondrial metabolism and ROS production were not significantly affected by the treatment either at chick or juvenile stages. Chicks in very small broods were smaller in size and had higher mitochondrial metabolic rates. The nest of rearing has a significant effect on nestling mitochondrial metabolism, yet variation in mitochondrial metabolism at the early-life stages are not associated with survival chances. The contribution of the rearing environment in determining offspring mitochondrial metabolism emphasizes the plasticity of mitochondrial metabolism in changing environments. Further studies would be needed to closely investigate what are the major environmental cues affecting the offspring mitochondrial metabolism during the growth period.

physiology↗

Can increased prenatal exposure to thyroid hormones alter physiology and behaviour in the long-term? Insights from an experimental study in Japanese quails.

Maternal thyroid hormones (triiodothyronine T3 and thyroxine T4) are important regulators of embryonic development and gene expression. While maternal thyroid disorders are known to cause developmental issues in humans, variation in maternal thyroid hormones in healthy mothers have also been found to correlate with infant and child phenotypes. This suggests a potential impact of maternal thyroid hormones on offspring phenotype in an eco-evolutionary context. In chickens, prenatal thyroid hormone treatment has been shown to influence embryonic gene expression and postnatal treatment to influence imprinting and learning. However, the potential long-term effects of maternal thyroid hormones on physiology and behaviour are unclear. This study aims to investigate the long-term effects of maternal thyroid hormones on behaviour, plasma thyroid hormone levels and brain gene expression using the Japanese quail (Coturnix japonica) as a model. Egg hormone levels were elevated by injecting unincubated eggs with either saline (control), T3, T4 or a mixture of T3 and T4. Social motivation, boldness and fearfulness to predators were tested shortly after hatching and as adults. Plasma thyroid hormone levels and pallial expression of thyroid hormone receptor A, type 2 deiodinase, and nuclear receptor coactivator 1 were measured in adulthood. We found no evidence that elevated thyroid hormone levels in eggs affected behaviour, plasma hormone levels, or gene expression. This is the first study examining the potential long-term effects of elevated maternal thyroid hormones within the natural range. Although we found no evidence of long-term effects, other traits may still be affected and remain to be studied.

physiology↗

From maternal glucocorticoid and thyroid hormones to epigenetic regulation of gene expression: an experimental study in a wild bird species

Offspring phenotype at birth is determined by its genotype and the prenatal environment including exposure to maternal hormones. Variation in both maternal glucocorticoids and thyroid hormones can affect offspring phenotype. However, the underlying molecular mechanisms shaping the offspring phenotype, especially those contributing to long-lasting effects, remain unclear. Epigenetic changes (such as DNA methylation) have been postulated as mediators of long-lasting effects of early-life environment. In this study, we determined the effects of elevated prenatal glucocorticoid and thyroid hormones on handling stress response (breath rate), DNA methylation and gene expression of glucocorticoid receptor (GCR) and thyroid hormone receptor (THR) in great tit (Parus major). Eggs were injected before incubation onset with corticosterone (main avian glucocorticoid) and/or thyroid hormones (thyroxine and triiodothyronine) to simulate variation in maternal hormone deposition. Breath rate during handling and gene expression of GCR and THR were evaluated 14 days after hatching. Methylation status of GCR and THR genes were analyzed from the longitudinal blood samples taken 7 and 14 days after hatching, as well as in the following autumn. Elevated prenatal corticosterone level significantly increased the breath rate during handling, indicating enhanced stress response and/or metabolism. Prenatal corticosterone manipulation had CpG-site-specific effects on DNA methylation at the GCR putative promoter region, while it did not significantly affect GCR gene expression. GCR expression was negatively associated with earlier hatching date and chick size. THR methylation or expression did not exhibit any significant relationship with the hormonal treatments or the examined covariates, suggesting that TH signaling may be more robust due to its crucial role in development. This study supports the view that maternal corticosterone may influence offspring metabolism and stress response via epigenetic alterations, yet their possible adaptive role in optimizing offspring phenotype to the prevailing conditions, context-dependency, and the underlying molecular interplay needs further research.

evolutionary biology↗

The CRL4B E3 ligase regulates mitosis by recruiting phospho-specific DCAFs

The cullin-4 paralogs CUL4A and CUL4B assemble E3 ubiquitin ligase complexes regulating multiple chromatin-associated cellular functions. Although they are structurally similar, we found that the unique N-terminal extension of CUL4B is heavily phosphorylated during mitosis, and the phosphorylation pattern is perturbed in the CUL4B-P50L mutation causing X-linked intellectual disability (XLID). Phenotypic characterization and mutational analysis revealed that CUL4B phosphorylation is required for efficient progression through mitosis, controlling spindle positioning and cortical tension. Interestingly, while CUL4B phosphorylation triggers chromatin exclusion, it critically promotes binding to actin regulators and two previously unrecognized, CUL4B-specific DCAFs, LIS1 and WDR1. Indeed, co-immunoprecipitation experiments and biochemical analysis revealed that LIS1 and WDR1 interact with DDB1, but their binding requires the phosphorylated N-terminal domain of CUL4B. Together, our study uncovers previously unrecognized DCAFs relevant for mitosis and brain development that specifically bind CUL4B, but not the CUL4B-P50L patient mutant, by a phosphorylation-dependent mechanism.

cell biology↗

Associations between brood size, gut microbiome diversity and survival in great tit (Parus major) nestlings

BackgroundThe gut microbiome forms at an early stage, yet data on the environmental factors influencing the development of wild avian microbiomes is limited. As the gut microbiome is a vital part of organismal health, it is important to understand how it may connect to host performance. The early studies with wild gut microbiome have shown that the rearing environment may be of importance in gut microbiome formation, yet the results vary across taxa, and the effects of specific environmental factors have not been characterized. Here, wild great tit (Parus major) broods were manipulated to either reduce or enlarge the original brood soon after hatching. We investigated if brood size was associated with nestling bacterial gut microbiome, and whether gut microbiome diversity predicted survival. Fecal samples were collected at mid-nestling stage and sequenced with the 16S rRNA gene amplicon sequencing, and nestling growth and survival were measured. ResultsGut microbiome diversity showed high variation between individuals, but this variation was not significantly explained by brood size or body mass. Additionally, we did not find a significant effect of brood size on body mass or gut microbiome composition. We also demonstrated that early handling had no impact on nestling performance or gut microbiome. Furthermore, we found no significant association between gut microbiome diversity and short-term (survival to fledging) or mid-term (apparent juvenile) survival. ConclusionsWe found no clear association between early-life environment, offspring condition and gut microbiome. This suggests that brood size is not a significantly contributing factor to great tit nestling condition, and that other environmental and genetic factors may be more strongly linked to offspring condition and gut microbiome. Future studies should expand into other early-life environmental factors e.g., diet composition and quality, and parental influences.

ecology↗

Prenatal thyroid hormones accelerate postnatal growth and telomere shortening in wild great tits

Early-life environment is known to affect later-life health and disease, which could be mediated by the early-life programming of telomere length, a key hallmark of ageing. According to the fetal programming of telomere biology hypothesis, variation in prenatal exposure to hormones is likely to influence telomere length. Yet the contribution of key metabolic hormones, i.e. thyroid hormones (THs), has been largely ignored. We recently showed that in contrast to predictions, exposure to elevated prenatal THs increased postnatal telomere length in wild collared flycatchers, but the generality of such effect, its underlying proximate mechanisms and consequences on survival have not been investigated. We therefore conducted a comprehensive study evaluating the impact of THs on potential drivers of telomere dynamics (growth, post-natal THs, mitochondria and oxidative stress), telomere length and medium-term survival using wild great tits as a model system. While prenatal THs did not significantly affect telomere length a week after hatching (i.e. day 7), they influenced postnatal telomere shortening (i.e. shorter telomeres at day 14 and the following winter) but not apparent survival. Circulating THs, mitochondrial density or oxidative stress biomarkers were not significantly influenced, whereas TH-supplemented group showed accelerated growth, which may explain the observed delayed effect on telomeres. We discuss several alternative hypotheses that may explain the contrast with our previous findings in flycatchers. Given that shorter telomeres in early life tend to be carried until adulthood and are often associated with decreased survival prospects, the effects of prenatal THs on telomeres may have long-lasting effects on senescence.

evolutionary biology↗

Human blood contains circulating cell-free mitochondria, but are they really functional?

Dache et al. (2020, FASEB J. 15, e2002338-15) recently reported the presence of respiratory-competent cell-free mitochondria in human blood (up to 3.7 x 106 per mL of blood), providing exciting perspectives on the potential role of these extra-cellular mitochondria. While their evidence for the presence of cell-free mitochondria in human blood is compelling, their conclusion that these cell-free mitochondria are respiratory-competent or functional has to be re-evaluated. To this end, we evaluated the functionality of cell-free mitochondria in human blood using high-resolution respirometry and mitochondria extracted from platelets of the same blood samples as positive controls. While cell-free mitochondria were present in human plasma (i.e. significant MitoTracker Green Fluorescence and complex IV activity), there was no evidence suggesting that their mitochondrial electron transport system (ETS) was functional (i.e. respiration rate not significantly different from 0; no significant responses to ADP, uncoupler or mitochondrial inhibitors oligomycin and antimycin A). Yet, in vitro complex IV activity was detectable and even slightly higher than levels found in mitochondria extracted from platelets, suggesting that cell-free mitochondria in human blood are likely to only retain a non-functional part of the electron transport system. Despite being unlikely to be fully functional in the narrow-sense (i.e. capable of oxidative phosphorylation), circulating cell-free mitochondria may have significant physiological roles that remain to be elucidated.

physiology↗

Born to be young: prenatal thyroid hormones increase early-life telomere length

Prenatal environmental conditions can have lifelong consequences on health and aging. The underlying mechanisms remain nonetheless little understood. Thyroid hormones (THs) are important regulators of embryogenesis transferred from the mother to the embryo. In an avian model, we manipulated embryo exposure to maternal THs through egg injection and investigated the consequences on postnatal growth and aging. We first report that mitochondrial DNA (mtDNA) copy number and telomere length significantly decrease from early-life to late adulthood, thus confirming that these two molecular markers are hallmarks of aging in our wild bird model. The experimental elevation of prenatal THs levels had a transient positive effect on postnatal growth. Elevated prenatal THs had no effect on mtDNA copy number but significantly increased telomere length both soon after birth and at the end of the growth period (equivalent to offsetting ca. 4 years of post-growth telomere shortening). These findings suggest that prenatal THs have a key role in setting the biological age at birth, and thus might influence longevity.

physiology↗

Ageing before birth: pace and stability of prenatal growth affect telomere dynamics

Pre-natal effects on telomere length are increasingly recognized as a potential contributor to the developmental origin of health and adult diseases. While it is becoming clear that telomere length is strongly influenced by pre-natal conditions, the factors affecting telomere dynamics during embryogenesis remain poorly understood. We manipulated both the pace and stability of prenatal growth using incubation temperature in Japanese quail and investigated the impact on telomere dynamics from embryogenesis to adulthood, along with potential drivers of telomere shortening such as oxidative damage and prenatal glucocorticoid levels. Telomere length was not affected by these manipulations for the first 75% of prenatal development, but was reduced at hatching in response to both experimentally induced fast embryo growth and growth instability. These early-life effects on telomere length persisted until adulthood. The effect of developmental instability on telomere length at hatching was potentially mediated by an increased secretion of glucocorticoid hormones during development. Both the pace and the stability of prenatal growth appear as key factors determining telomere length and dynamics up to adulthood, with fast and unstable embryo growth leading to short telomeres with the potential for adverse associated outcomes in terms of reduced longevity and greater risk of disease.

physiology↗