Directional selection, not the direction of selection, affects telomere length and copy number at ribosomal RNA loci.

Many fisheries exert directional selection on traits such as body size and growth rate. Whether directional selection impacts regions of the genome associated with traits related to growth is unknown. To address this issue, we characterised copy number variation in three regions of the genome associated with cell division, (1) telomeric DNA, (2) loci transcribed as ribosomal RNA (rDNA), and (3) mitochondrial DNA (mtDNA), in three selection lines of zebrafish reared at three temperatures (22 °C, 28 °C, and 34 °C). Selection lines differed in (1) the direction of selection (two lines experienced directional selection for large or small body size) and (2) whether they experienced any directional selection itself. Lines that had experienced directional selection were smaller, had lower growth rate, shorter telomeres, and lower rDNA copy number than the line that experiencing no directional selection. Neither telomere length nor rDNA copy number were affected by temperature. In contrast, mtDNA content increased at elevated temperature but did not differ among selection lines. Though directional selection impacts rDNA and telomere length, direction of such selection did not matter, whereas mtDNA acts as a stress marker for temperature. Future work should examine the consequences of these genomic changes in natural fish stocks.

Does size-selective harvesting erode adaptive potential to thermal stress?

Overharvesting is a serious threat to many fish populations. High mortality and directional selection on body size can cause evolutionary change in exploited populations via selection for a specific phenotype and a potential reduction in phenotypic diversity. Whether the loss of phenotypic diversity that accompanies directional selection impairs response to environmental stress is not known. To address this question, we exposed three zebrafish selection lines to thermal stress. Two lines had experienced directional selection for (1) large and (2) small body size, and one was (3) subject to random removal of individuals with respect to body size (i.e. line with no directional selection). Selection lines were exposed to three temperatures (elevated, 34°C; ambient, 28°C; low, 22°C) to determine the response to an environmental stressor (thermal stress). We assessed differences among selection lines in their life history (growth and reproduction), physiological traits (metabolic rate and critical thermal max) and behaviour (activity and feeding behaviour) when reared at different temperatures. Lines experiencing directional selection (i.e. size selected) showed reduced growth rate and a shift in average phenotype in response to lower or elevated thermal stress compared with fish from the random-selected line. Our data indicate that populations exposed to directional selection can have a more limited capacity to respond to thermal stress compared with fish that experience a comparable reduction in population size (but without directional selection). Future studies should aim to understand the impacts of environmental stressors on natural fish stocks.

Parental cardiorespiratory fitness influences early life energetics and metabolic health.

High cardiorespiratory fitness (CRF) is associated with a reduced risk of metabolic disease and is linked to superior mitochondrial respiratory function. This study investigated how intrinsic CRF affects bioenergetics and metabolic health in adulthood and early life. Adult rats selectively bred for low and high running capacity [low capacity runners (LCR) and high capacity runners (HCR), respectively] underwent metabolic phenotyping before mating. Weanlings were evaluated at 4-6 wk of age, and whole body energetics and behavior were assessed using metabolic cages. Mitochondrial respiratory function was assessed in permeabilized tissues through high-resolution respirometry. Proteomic signatures of adult and weanling tissues were determined using mass spectrometry. The adult HCR group exhibited lower body mass, improved glucose tolerance, and greater physical activity compared with the LCR group. The adult HCR group demonstrated higher mitochondrial respiratory capacities in the soleus and heart compared with the adult LCR group, which coincided with a greater abundance of proteins involved in lipid catabolism. HCR and LCR weanlings had similar body mass, but HCR weanlings displayed reduced adiposity. In addition, HCR weanlings exhibited better glucose tolerance and higher physical activity levels than LCR weanlings. Higher respiratory capacities were observed in the soleus, heart, and liver tissues of HCR weanlings compared with LCR weanlings, which were not owed to greater mitochondrial content. Proteomic analyses indicated a greater potential for lipid oxidation in the contractile muscles of HCR weanlings. In conclusion, offspring born to parents with high CRF possess an enhanced capacity for lipid catabolism and oxidative phosphorylation, thereby influencing metabolic health. These findings highlight that intrinsic CRF shapes the bioenergetic phenotype with implications for metabolic resilience in early life.NEW & NOTEWORTHY Inherited cardiorespiratory fitness (CRF) influences early life bioenergetics and metabolic health. Higher intrinsic CRF was associated with reduced adiposity and improved glucose tolerance in early life. This metabolic phenotype was accompanied by greater mitochondrial respiratory capacity in skeletal muscle, heart, and liver tissue. Proteomic profiling of these three tissues further revealed potential mechanisms linking inherited CRF to early life metabolism.

Elevated shear rate-induced by exercise increases eNOS ser1177 but not PECAM-1 Tyr713 phosphorylation in human conduit artery endothelial cells.

Although evidence demonstrates the fundamental role of shear stress in vascular health, predominantly through the release of nitric oxide (NO), the mechanisms by which endothelial cells (EC)s sense and transduce shear are poorly understood. In cultured ECs tyrosine phosphorylation of PECAM-1 has been shown to activate eNOS in response to shear stress. However, in the human skeletal muscle microcirculation PECAM-1 was not activated in response to exercise or passive leg movement. Given this contradiction, this study aimed to assess the effect of exercise on conduit artery PECAM-1 and eNOS activation in humans. Eleven males were randomised to two groups; 30 min of handgrip exercise (n = 6), or a time-control group (n = 5). Protein content of eNOS and PECAM-1, alongside eNOS Ser1177 and PECAM-1 Tyr713 phosphorylation were assessed in ECs obtained from the radial artery pre- and post-intervention. Handgrip exercise resulted in a 5-fold increase in mean shear rate in the exercise group, with no change in the control group (group*time, P < 0.001). There was a 54% increase in eNOS Ser1177 phosphorylation in the exercise group, when compared to control group (group*time, P = 0.016), but no change was reported in PECAM-1 Tyr713 phosphorylation in either group (group*time, P > 0.05). eNOS and PECAM-1 protein content were unchanged (group*time, P > 0.05). Our data show that exercise-induced elevations in conduit artery shear rate increase eNOS Ser1177 phosphorylation but not PECAM-1 Tyr713 phosphorylation. This suggests PECAM-1 phosphorylation may not be involved in the vascular response to acute but prolonged elevations in exercise-induced shear rate in conduit arteries of healthy, active men.

A modest change in housing temperature alters whole body energy expenditure and adipocyte thermogenic capacity in mice.

Typical vivarium temperatures (20-26°C) induce facultative thermogenesis in mice, a process attributable in part to uncoupling protein-1 (UCP1). The impact of modest changes in housing temperature on whole body and adipose tissue energetics in mice remains unclear. Here, we determined the effects of transitioning mice from 24°C to 30°C on total energy expenditure and adipose tissue protein signatures. C57BL/6J mice were housed at 24°C for 2 wk and then either remained at 24°C (n = 16/group, 8M/8F) or were transitioned to 30°C (n = 16/group, 8M/8F) for 4 wk. Total energy expenditure and its components were determined by indirect calorimetry. Interscapular brown adipose tissue (iBAT) and inguinal white adipose tissue (iWAT) proteins were quantified by Western blot and quantitative proteomics. Transitioning from 24°C to 30°C reduced total energy expenditure in both male (-25%) and female (-16%) mice, which was attributable to lower basal energy expenditure in males (-36%) and females (-40%). Total iBAT UCP1 protein content was 50% lower at 30°C compared with 24°C, whereas iWAT UCP1 protein content was similar between conditions. iBAT UCP1 protein content remained 20-fold greater than iWAT at 30°C. In iBAT and iWAT, 183 and 41 proteins were differentially expressed between 24°C and 30°C, respectively. iWAT proteins (257) differentially expressed between sexes at 30°C were not differentially expressed at 24°C. Thus, 30°C housing lowers total energy expenditure of mice when compared with an ambient temperature (24°C) that falls within the National Research Council's guidelines for housing laboratory mice. Lower iBAT UCP1 content accompanied chronic housing at 30°C. Furthermore, housing temperature influences sexual dimorphism in the iWAT proteome. These data have implications regarding the optimization of preclinical models of human disease.NEW & NOTEWORTHY Housing mice at 30°C reduced the basal and total energy expenditure compared with 24°C, which was accompanied by a reduction in brown adipose tissue UCP1 content. Proteomic profiling demonstrated the brown adipose tissue and white adipose tissue proteomes were largely influenced by housing temperature and sex, respectively. Therefore, 30°C housing revealed sexual dimorphism in the white adipose tissue proteome that was largely absent in animals housed at 24°C.

(-)-Epicatechin Alters Reactive Oxygen and Nitrogen Species Production Independent of Mitochondrial Respiration in Human Vascular Endothelial Cells.

INTRODUCTION: Vascular endothelial dysfunction is characterised by lowered nitric oxide (NO) bioavailability, which may be explained by increased production of reactive oxygen species (ROS), mitochondrial dysfunction, and altered cell signalling. (-)-Epicatechin (EPI) has proven effective in the context of vascular endothelial dysfunction, but the underlying mechanisms associated with EPI's effects remain unclear. Objective(s). Our aim was to investigate whether EPI impacts reactive oxygen and nitrogen species (RONS) production and mitochondrial function of human vascular endothelial cells (HUVECs). We hypothesised that EPI would attenuate ROS production, increase NO bioavailability, and enhance indices of mitochondrial function. METHODS: HUVECs were treated with EPI (0-20 µM) for up to 48 h. Mitochondrial and cellular ROS were measured in the absence and presence of antimycin A (AA), an inhibitor of the mitochondrial electron transport protein complex III, favouring ROS production. Genes associated with mitochondrial remodelling and the antioxidant response were quantified by RT-qPCR. Mitochondrial bioenergetics were assessed by respirometry and signalling responses determined by western blotting. RESULTS: Mitochondrial superoxide production without AA was increased 32% and decreased 53% after 5 and 10 µM EPI treatment vs. CTRL (P < 0.001). With AA, only 10 µM EPI increased mitochondrial superoxide production vs. CTRL (25%, P < 0.001). NO bioavailability was increased by 45% with 10 µM EPI vs. CTRL (P = 0.010). However, EPI did not impact mitochondrial respiration. NRF2 mRNA expression was increased 1.5- and 1.6-fold with 5 and 10 µM EPI over 48 h vs. CTRL (P = 0.015 and P = 0.001, respectively). Finally, EPI transiently enhanced ERK1/2 phosphorylation (2.9 and 3.2-fold over 15 min and 1 h vs. 0 h, respectively; P = 0.035 and P = 0.011). Conclusion(s). EPI dose-dependently alters RONS production of HUVECs but does not impact mitochondrial respiration. The induction of NRF2 mRNA expression with EPI might relate to enhanced ERK1/2 signalling, rather than RONS production. In humans, EPI may improve vascular endothelial dysfunction via alteration of RONS and activation of cell signalling.

Measuring Daphnia life history in the wild: The efficacy of individual field cages.

Life-history studies are often conducted in a laboratory environment where it is easy to assay individual animals. However, factors such as temperature, photoperiod, and nutrition vary greatly between laboratory and field environments, making it difficult to compare results. Consequently, there is a need to study individual life histories in the field, but this is currently difficult in systems such as Daphnia where it is not possible to mark and track individual animals. Here, we present a proof of principle study showing that field cages are a reliable method for collecting individual-level life-history data in Daphnia magna. As a first step, we compared the life history of paired animals reared outside and inside cages to test the hypothesis that cages allow free flow of algal food resources. We then used a seminatural mesocosm setting to compare the performance of individual field cages versus glass jars refilled with mesocosm water each day. We found that cages did not inhibit food flow and that differences in life histories between three clones detected in the jar assays were also detectable using the much less labor-intensive field cages. We conclude that field cages are a feasible approach for collecting individual-level life-history data in systems such as Daphnia where individual animals cannot be marked and tracked.

Cocoa-flavanols enhance moderate-intensity pulmonary [Formula: see text] kinetics but not exercise tolerance in sedentary middle-aged adults.

INTRODUCTION: Cocoa flavanols (CF) may exert health benefits through their potent vasodilatory effects, which are perpetuated by elevations in nitric oxide (NO) bioavailability. These vasodilatory effects may contribute to improved delivery of blood and oxygen (O2) to exercising muscle. PURPOSE: Therefore, the objective of this study was to examine how CF supplementation impacts pulmonary O2 uptake ([Formula: see text]) kinetics and exercise tolerance in sedentary middle-aged adults. METHODS: We employed a double-blind cross-over, placebo-controlled design whereby 17 participants (11 male, 6 female; mean ± SD, 45 ± 6 years) randomly received either 7 days of daily CF (400 mg) or placebo (PL) supplementation. On day 7, participants completed a series of 'step' moderate- and severe-intensity exercise tests for the determination of [Formula: see text] kinetics. RESULTS: During moderate-intensity exercise, the time constant of the phase II [Formula: see text] kinetics ([Formula: see text]) was decreased by 15% in CF as compared to PL (mean ± SD; PL 40 ± 12 s vs. CF 34 ± 9 s, P = 0.019), with no differences in the amplitude of [Formula: see text] (A[Formula: see text]; PL 0.77 ± 0.32 l min-1 vs. CF 0.79 ± 0.34 l min-1, P = 0.263). However, during severe-intensity exercise, [Formula: see text], the amplitude of the slow component ([Formula: see text]) and exercise tolerance (PL 435 ± 58 s vs. CF 424 ± 47 s, P = 0.480) were unchanged between conditions. CONCLUSION: Our data show that acute CF supplementation enhanced [Formula: see text] kinetics during moderate-, but not severe-intensity exercise in middle-aged participants. These novel effects of CFs, in this demographic, may contribute to improved tolerance of moderate-activity physical activities, which appear commonly present in daily life. TRIAL REGISTRATION: Registered under ClinicalTrials.gov Identifier no. NCT04370353, 30/04/20 retrospectively registered.

Temperature and clone-dependent effects of microplastics on immunity and life history in Daphnia magna.

Microplastic (MP) pollution is potentially a major threat to many aquatic organisms. Yet we currently know very little about the mechanisms responsible for the effects of small MPs on phenotypes, and the extent to which effects of MPs are modified by genetic and environmental factors. Using a multivariate approach, we studied the effects of 500 nm polystyrene microspheres on the life history and immunity of eight clones of the freshwater cladoceran Daphnia magna reared at two temperatures (18 °C/24 °C). MP exposure altered multivariate phenotypes in half of the clones we studied but had no effect on others. In the clones that were affected, individuals exposed to MPs had smaller offspring at both temperatures, and more offspring at high temperature. Differences in response to MP exposure were unrelated to differences in particle uptake, but were instead linked to an upregulation of haemocytes, particularly at high temperature. The clone-specific, context-dependent nature of our results demonstrates the importance of incorporating genetic variation and environmental context into assessments of the impact of plastic particle exposure. Our results identify immunity as an important mechanism underpinning genetically variable responses to MP pollution and may have major implications for predicting consequences of MP pollution.

The effects of elevated CO2 on shell properties and susceptibility to predation in mussels Mytilus edulis.

For many species, ocean acidification (OA) is having negative physiological consequences on their fitness and resilience to environmental change, but less is known about the ecosystem effects of these changes. Here, we assess how OA conditions predicted for 2100 affects the biological functioning of an important habitat-forming species Mytilus edulis and its susceptibility to predation by a key predator, the gastropod Nucella lapillus. Change in three physiological parameters in Mytilus were assessed: (1) shell thickness and cross-sectional surface area, (2) body volume and (3) feeding rate, as well as susceptibility to predation by N. lapillus. Shell thickness and cross-section area, body volume and feeding rate of Mytilus all reduced under OA conditions indicating compromised fitness. Predation risk increased by ∼26% under OA, suggesting increased susceptibility of mussels to predation and/or altered predator foraging behaviour. Notably, predation of large Mytilus - that were largely free from predation under control conditions - increased by more than 8x under OA, suggesting that body size was no longer a refuge. Our results suggest OA will impact upon ecosystem structure and functioning and the continued provision of ecosystem services associated with Mytilus reefs and the communities associated with them.

Can false memory for critical lures occur without conscious awareness of list words?

We examined whether the DRM false memory effect can occur when list words are presented below the perceptual identification threshold. In four experiments, subjects showed robust veridical memory for studied words and false memory for critical lures when masked list words were presented at exposure durations of 43 ms per word. Shortening the exposure duration to 29 ms virtually eliminated veridical recognition of studied words and completely eliminated false recognition of critical lures. Subjective visibility ratings in Experiments 3a and 3b support the assumption that words presented at 29 ms were subliminal for most participants, but were occasionally experienced with partial awareness by participants with higher perceptual awareness. Our results indicate that a false memory effect does not occur in the absence of conscious awareness of list words, but it does occur when word stimuli are presented at an intermediate level of visibility.

Tunable cw UV laser with <35 kHz absolute frequency instability for precision spectroscopy of Sr Rydberg states.

We present a solid-state laser system that generates over 200 mW of continuous-wave, narrowband light, tunable from 316.3 nm - 317.7 nm and 318.0 nm - 319.3 nm. The laser is based on commercially available fiber amplifiers and optical frequency doubling technology, along with sum frequency generation in a periodically poled stoichiometric lithium tantalate crystal. The laser frequency is stabilized to an atomic-referenced high finesse optical transfer cavity. Using a GPS-referenced optical frequency comb we measure a long term frequency instability of < 35 kHz for timescales between 10(-3) s and 10(3) s. As an application we perform spectroscopy of Sr Rydberg states from n = 37 - 81, demonstrating mode-hop-free scans of 24 GHz. In a cold atomic sample we measure Doppler-limited linewidths of 350 kHz.

Physical and biogeochemical modulation of ocean acidification in the central North Pacific.

Atmospheric carbon dioxide (CO(2)) is increasing at an accelerating rate, primarily due to fossil fuel combustion and land use change. A substantial fraction of anthropogenic CO(2) emissions is absorbed by the oceans, resulting in a reduction of seawater pH. Continued acidification may over time have profound effects on marine biota and biogeochemical cycles. Although the physical and chemical basis for ocean acidification is well understood, there exist few field data of sufficient duration, resolution, and accuracy to document the acidification rate and to elucidate the factors governing its variability. Here we report the results of nearly 20 years of time-series measurements of seawater pH and associated parameters at Station ALOHA in the central North Pacific Ocean near Hawaii. We document a significant long-term decreasing trend of -0.0019 +/- 0.0002 y(-1) in surface pH, which is indistinguishable from the rate of acidification expected from equilibration with the atmosphere. Superimposed upon this trend is a strong seasonal pH cycle driven by temperature, mixing, and net photosynthetic CO(2) assimilation. We also observe substantial interannual variability in surface pH, influenced by climate-induced fluctuations in upper ocean stability. Below the mixed layer, we find that the change in acidification is enhanced within distinct subsurface strata. These zones are influenced by remote water mass formation and intrusion, biological carbon remineralization, or both. We suggest that physical and biogeochemical processes alter the acidification rate with depth and time and must therefore be given due consideration when designing and interpreting ocean pH monitoring efforts and predictive models.

Climate-driven changes to the atmospheric CO2 sink in the subtropical North Pacific Ocean.

The oceans represent a significant sink for atmospheric carbon dioxide. Variability in the strength of this sink occurs on interannual timescales, as a result of regional and basin-scale changes in the physical and biological parameters that control the flux of this greenhouse gas into and out of the surface mixed layer. Here we analyse a 13-year time series of oceanic carbon dioxide measurements from station ALOHA in the subtropical North Pacific Ocean near Hawaii, and find a significant decrease in the strength of the carbon dioxide sink over the period 1989-2001. We show that much of this reduction in sink strength can be attributed to an increase in the partial pressure of surface ocean carbon dioxide caused by excess evaporation and the accompanying concentration of solutes in the water mass. Our results suggest that carbon dioxide uptake by ocean waters can be strongly influenced by changes in regional precipitation and evaporation patterns brought on by climate variability.