CO2 Sensitivity Research Articles

Activation of Phox2b-Expressing Neurons in the Nucleus Tractus Solitarii Drives Breathing in Mice.

The nucleus tractus solitarii (NTS) is implicated in the control of breathing, but the neuronal phenotype and circuit mechanism involved in such a physiological function remain incompletely understood. This study focused on the respiratory role of paired-like homeobox 2b gene (Phox2b)-expressing NTS neurons and sought to determine whether selective stimulation of this set of neurons activates breathing in male mice. A Cre-dependent vector encoding a Gq-coupled human M3 muscarinic receptor (hM3Dq) was microinjected into the NTS of Phox2b-Cre transgenic mice. The hM3Dq-transduced neurons were pharmacologically activated in conscious mice while respiratory effects were measured by plethysmography. We demonstrate that chemogenetic stimulation of Phox2b-expressing NTS neurons significantly increased baseline minute volume via an increase in respiratory frequency rather than tidal volume. Chemogenetic stimulation also synergized with moderate CO2 stimulation to enhance pulmonary ventilatory response. Selective ablation of Phox2b-expressing NTS neurons notably attenuated a hypercapnic ventilatory response. Moreover, histological evidence revealed that stimulation of Phox2b-expressing NTS neurons increased neuronal activity of the preBötzinger complex. Finally, we presented the neuroanatomical evidence of direct projection of Phox2b-expressing NTS neurons to putative respiratory central pattern generator. Overall, these findings suggest that selective activation of Phox2b-expressing NTS neurons potentiates baseline pulmonary ventilation via an excitatory drive to respiratory central pattern generator and this group of neurons is also required for the hypercapnic ventilatory response.SIGNIFICANCE STATEMENT The nucleus tractus solitarii (NTS) has been implicated in the control of breathing. The paired-like homeobox 2b gene (Phox2b) is the disease-defining gene for congenital central hypoventilation syndrome and is restrictively present in brainstem nucleus, including the NTS. Using a chemogenetic approach, we demonstrate herein that selective stimulation of Phox2b-expressing NTS neurons vigorously potentiates baseline pulmonary ventilation via an excitatory drive to respiratory central pattern generator in rodents. Genetic ablation of these neurons attenuates the hypercapnic ventilatory response. We also suggest that a fraction of Phox2b-expressing neurons exhibit CO2 sensitivity and presumably function as central respiratory chemoreceptors. The methodology is expected to provide a future applicability to the patients with sleep-related hypoventilation or apnea.

High sensitivity of a keystone forage fish to elevated CO2 and temperature.

Sand lances of the genus Ammodytes are keystone forage fish in coastal ecosystems across the northern hemisphere. Because they directly support populations of higher trophic organisms such as whales, seabirds or tuna, the current lack of empirical data and, therefore, understanding about the climate sensitivity of sand lances represent a serious knowledge gap. Sand lances could be particularly susceptible to ocean warming and acidification because, in contrast to other tested fish species, they reproduce during boreal winter months, and their offspring develop slowly under relatively low and stable pCO2 conditions. Over the course of 2 years, we conducted factorial pCO2 × temperature exposure experiments on offspring of the northern sand lance Ammodytes dubius, a key forage species on the northwest Atlantic shelf. Wild, spawning-ripe adults were collected from Stellwagen Bank National Marine Sanctuary (Cape Cod, USA), and fertilized embryos were reared at three pCO2 conditions (400, 1000 and 2100 μatm) crossed with three temperatures (5, 7 and 10 ˚C). Exposure to future pCO2 conditions consistently resulted in severely reduced embryo survival. Sensitivity to elevated pCO2 was highest at 10 ˚C, resulting in up to an 89% reduction in hatching success between control and predicted end-of-century pCO2 conditions. Moreover, elevated pCO2 conditions delayed hatching, reduced remaining endogenous energy reserves at hatch and reduced embryonic growth. Our results suggest that the northern sand lance is exceptionally CO2-sensitive compared to other fish species. Whether other sand lance species with similar life history characteristics are equally CO2-sensitive is currently unknown. But the possibility is a conservation concern, because many boreal shelf ecosystems rely on sand lances and might therefore be more vulnerable to climate change than currently recognized. Our findings indicate that life history, spawning habitat, phenology and developmental rates mediate the divergent early life CO2 sensitivities among fish species.

Robust quantification of fish early life CO2 sensitivities via serial experimentation.

Despite the remarkable expansion of laboratory studies, robust estimates of single species CO2 sensitivities remain largely elusive. We conducted a meta-analysis of 20 CO2 exposure experiments conducted over 6 years on offspring of wild Atlantic silversides (Menidia menidia) to robustly constrain CO2 effects on early life survival and growth. We conclude that early stages of this species are generally tolerant to CO2 levels of approximately 2000 µatm, likely because they already experience these conditions on diel to seasonal timescales. Still, high CO2 conditions measurably reduced fitness in this species by significantly decreasing average embryo survival (-9%) and embryo+larval survival (-13%). Survival traits had much larger coefficients of variation (greater than 30%) than larval length or growth (3-11%). CO2 sensitivities varied seasonally and were highest at the beginning and end of the species' spawning season (April-July), likely due to the combined effects of transgenerational plasticity and maternal provisioning. Our analyses suggest that serial experimentation is a powerful, yet underused tool for robustly estimating small but true CO2 effects in fish early life stages.

Comparison of methods of chemical loop gain measurement during tidal ventilation in awake healthy subjects.

The loop gain (LG) is defined as the ratio of a ventilatory response over the perturbation in ventilation, and it is used to analyze ventilatory control stability. The LG can be derived from minute ventilation (V̇e), end-tidal Pco2 ([Formula: see text]), and end-tidal Po2 ([Formula: see text]) values. Several methods of LG assessment have been developed, which have never been compared. We evaluated the computability, the short-term repeatability, and the agreement of six published (or slightly modified) models for LG determination. These models included three unconstrained autoregressive models, univariate (V̇e), bivariate (V̇e, [Formula: see text]), and trivariate (V̇e, [Formula: see text], and [Formula: see text]), and three analytical transfer function constrained models based on V̇e, V̇e and CO2-sensitivity, and V̇e and central and peripheral CO2 sensitivities, respectively. The models were tested with tidal breathing data in 37 awake healthy subjects (median age 35 yr; 23 women, 14 men). Modeling failed in 11, 0, and 0 subjects for the three unconstrained models, respectively, and 4, 1, and 9 subjects for the three constrained models, respectively. Bland and Altman analyses of the LG values in the medium frequency range of two separate recordings demonstrated good repeatability for four models, excluding univariate and trivariate unconstrained models. The four repeatable models gave LG values that were in agreement (medium frequency LG, median 0.100-0.210), although the constrained model based on V̇e systematically overestimated LG values. The variances explained by these models were ∼20%. In conclusion, model-based analyses of tidal breathing were performed with different approaches that gave comparable results for chemical LG and explained variance.NEW & NOTEWORTHY Several methods of chemical loop gain measurement have been published but never compared. We show that a better repeatability is obtained with analytical constrained models compared with autoregressive unconstrained models and that the repeatable models gave comparable results of loop gain, even if the calculation based on ventilation-only recording gave higher values than those obtained with both ventilation and end-tidal Pco2 recording. The explained variance of ventilation was similar whatever the model.

Is the Last Glacial Maximum a reverse analog for future hydroclimate changes in the Americas?

Future hydroclimate change is expected to generally follow a wet-get-wetter, dry-get-drier (WWDD) pattern, yet key uncertainties remain regionally and over land. It has been previously hypothesized that lake levels of the Last Glacial Maximum (LGM) could map a reverse analog to future hydroclimate changes due to reduction of CO2 levels at this time. Potential complications to this approach include, however, the confounding effects of factors such as the Laurentide Ice Sheet and lake evaporation changes. Using the ensemble output of six coupled climate models, lake energy and water balance models, an atmospheric moisture budget analysis, and additional CO2 sensitivity experiments, we assess the effectiveness of the LGM as a reverse analog for future hydroclimate changes for a transect from the drylands of North America to southern South America. The model ensemble successfully simulates the general pattern of lower tropical lake levels and higher extratropical lake levels at LGM, matching 82% of the lake proxy records. The greatest model-data mismatch occurs in tropical and extratropical South America, potentially as a result of underestimated changes in temperature and surface evaporation. Thermodynamic processes of the mean circulation best explain the direction of lake changes observed in the proxy record, particularly in the tropics and Pacific coasts of the extratropics, and produce a WWDD pattern. CO2 forcing alone cannot account for LGM lake level changes, however, as the enhanced cooling from the Laurentide ice sheet appears necessary to generate LGM dry anomalies in the tropics and to deepen anomalies in the extratropics. LGM performance as a reverse analog is regionally dependent as anti-correlation between LGM and future P − E is not uniformly observed across the study domain.

Three Methods of Estimating Mesophyll Conductance Agree Regarding its CO₂ Sensitivity in the Rubisco-Limited Ci Range.

Whether the mesophyll conductance to CO2 movement (gm) within leaves of C3 plants changes with CO2 concentration remains a matter of debate, particularly at low CO2 concentrations. We tested for changes in gm over the range of sub-stomatal CO2 concentrations (Ci) for which Rubisco activity limited photosynthesis (A) in three plant species grown under the same conditions. Mesophyll conductance was estimated by three independent methods: the oxygen sensitivity of photosynthesis, variable J fluorescence combined with gas exchange, and the curvature of the Rubisco-limited A vs. Ci curve. The latter assay used a new method of rapidly obtaining data points at approximately every 3 μmol mol−1 for Rubisco-limited A vs. Ci curves, allowing separate estimates of curvature over limited Ci ranges. In two species, soybean and sunflower, no change in gm with Ci was detected using any of the three methods of estimating gm. In common bean measured under the same conditions as the other species, all three methods indicated large decreases in gm with increasing Ci. Therefore, change in gm with Ci in the Rubsico-limited region of A vs. Ci curves depended on the species, but not on the method of estimating gm.

You Better Repeat It: Complex CO2 × Temperature Effects in Atlantic Silverside Offspring Revealed by Serial Experimentation

Concurrent ocean warming and acidification demand experimental approaches that assess biological sensitivities to combined effects of these potential stressors. Here, we summarize five CO2 × temperature experiments on wild Atlantic silverside, Menidia menidia, offspring that were reared under factorial combinations of CO2 (nominal: 400, 2200, 4000, and 6000 µatm) and temperature (17, 20, 24, and 28 °C) to quantify the temperature-dependence of CO2 effects in early life growth and survival. Across experiments and temperature treatments, we found few significant CO2 effects on response traits. Survival effects were limited to a single experiment, where elevated CO2 exposure reduced embryo survival at 17 and 24 °C. Hatch length displayed CO2 × temperature interactions due largely to reduced hatch size at 24 °C in one experiment but increased length at 28 °C in another. We found no overall influence of CO2 on larval growth or survival to 9, 10, 15 and 13–22 days post-hatch, at 28, 24, 20, and 17 °C, respectively. Importantly, exposure to cooler (17 °C) and warmer (28 °C) than optimal rearing temperatures (24 °C) in this species did not appear to increase CO2 sensitivity. Repeated experimentation documented substantial inter- and intra-experiment variability, highlighting the need for experimental replication to more robustly constrain inherently variable responses. Taken together, these results demonstrate that the early life stages of this ecologically important forage fish appear largely tolerate to even extreme levels of CO2 across a broad thermal regime.

Thermal sensitivity of CO2 and CH4 emissions varies with streambed sediment properties

Globally, rivers and streams are important sources of carbon dioxide and methane, with small rivers contributing disproportionately relative to their size. Previous research on greenhouse gas (GHG) emissions from surface water lacks mechanistic understanding of contributions from streambed sediments. We hypothesise that streambeds, as known biogeochemical hotspots, significantly contribute to the production of GHGs. With global climate change, there is a pressing need to understand how increasing streambed temperatures will affect current and future GHG production. Current global estimates assume linear relationships between temperature and GHG emissions from surface water. Here we show non-linearity and threshold responses of streambed GHG production to warming. We reveal that temperature sensitivity varies with substrate (of variable grain size), organic matter (OM) content and geological origin. Our results confirm that streambeds, with their non-linear response to projected warming, are integral to estimating freshwater ecosystem contributions to current and future global GHG emissions.

A new oxadiazole-based dual-mode chemosensor: Colorimetric detection of Co2+ and fluorometric detection of Cu2+ with high selectivity and sensitivity

An asymmetrical oxadiazole derivative (Sensor 1) containing one pyridine and one formyl connecting to two benzene rings on both sides of oxadiazole unit was designed and synthesized. Sensor 1 displayed selective colorimetric sensing for Co2+ and Cu2+ ions via color changes to blue and yellow from colorless in CH3CN solutions, respectively. Sensor 1 not only exhibited an instantaneous naked-eye color change and but also could be used as a convenient test kit for recognition and detection of Co2+ ions. Sensor 1 presented high colorimetric sensitivity for Co2+ with low detection limit (3.92 × 10−6 mol/L). Sensor 1 also displayed “On-Off” fluorescent sensing for Cu2+ over a wide range of competitive metal ions and could accurately detect the existence of Cu2+ in water samples via the fluorescent quenching and color change. The fluorescence intensity of sensor 1 in CH3CN solution showed a linear response to Cu2+ in the wide concentration range of 0–40 μM which is beneficial for quantitative analysis. The studies on absorbance and fluorescence titration, 1H NMR spectrum and ESI-MS spectra confirmed that colorimetric sensing for Co2+ and fluorescent sensing for Cu2+ are ascribed to the complex formation with the stoichiometry of 1:1 for Co2+ and 1:2 for Cu2+.

Comparison of CO2 Flow Behavior through Intact Siltstone Sample under Tri-Axial Steady-State and Transient Flow Conditions

With its low viscosity properties, CO2 has much greater penetration capacity into micro-fractures, and therefore has more potential to create expanded and effective fractures in shales during the hydraulic fracturing process. However, the feasibility of this technique is dependent on the accurate prediction of formation flow characteristics, given the high leak-off of CO2 at deep depths. The aim of this study is therefore to understand the flow behavior of CO2 in deep shale plays. A series of tri-axial permeability tests was conducted under both steady-state and transient conditions. The test results show much lower permeability values for liquid CO2 than gaseous CO2, and the permeability under transient conditions is much lower than that under steady-state conditions, due to the combined effects of the reduced slip-flow effect under low pressures and the temperature variation influence under steady-state conditions. Under steady-state conditions, unstable flow behavior occurred at higher injection pressure (≥9 MPa) possibly due to the fine mineral particle migration and the deposition of small drikold particles, which indicates the serious error in permeability calculation under steady-state conditions. Importantly, a greater than 1 effective stress coefficient (χ) for permeability in tested siltstone was observed, confirming the greater sensitivity of CO2 to pore pressure than confining pressure.

Geographical CO2 sensitivity of phytoplankton correlates with ocean buffer capacity.

Accumulation of anthropogenic CO2 is significantly altering ocean chemistry. A range of biological impacts resulting from this oceanic CO2 accumulation are emerging, however, the mechanisms responsible for observed differential susceptibility between organisms and across environmental settings remain obscure. A primary consequence of increased oceanic CO2 uptake is a decrease in the carbonate system buffer capacity, which characterizes the system's chemical resilience to changes in CO2 , generating the potential for enhanced variability in pCO2 and the concentration of carbonate [ ], bicarbonate [ ], and protons [H+ ] in the future ocean. We conducted a meta-analysis of 17 shipboard manipulation experiments performed across three distinct geographical regions that encompassed a wide range of environmental conditions from European temperate seas to Arctic and Southern oceans. These data demonstrated a correlation between the magnitude of natural phytoplankton community biological responses to short-term CO2 changes and variability in the local buffer capacity across ocean basin scales. Specifically, short-term suppression of small phytoplankton (<10μm) net growth rates were consistently observed under enhanced pCO2 within experiments performed in regions with higher ambient buffer capacity. The results further highlight the relevance of phytoplankton cell size for the impacts of enhanced pCO2 in both the modern and future ocean. Specifically, cell size-related acclimation and adaptation to regional environmental variability, as characterized by buffer capacity, likely influences interactions between primary producers and carbonate chemistry over a range of spatio-temporal scales.

The effect of caffeine on the ventilatory response to hypercarbia in preterm infants.

BackgroundWe tested the hypotheses that caffeine therapy would increase the ventilatory response to hypercarbia in infants above the effect of maturation and those with a weaker ventilatory response to hypercarbia would be more likely to subsequently develop apnea that required treatment.MethodsInfants born at less than 34 weeks of gestation underwent a steady-state hypercarbic challenge using 0, 2, and 4% carbon dioxide soon after birth that was repeated at weekly intervals. The results of the initial study were compared between infants who did or did not subsequently develop apnea requiring treatment with caffeine.ResultsTwenty-six infants born at a median gestation of 32 (range 31-33) weeks were assessed. Caffeine administration was associated with an increase in CO2 sensitivity, and the mean increase was 15.3 (95% CI: 1-30) ml/kg/min/% CO2. Fourteen infants subsequently developed apnea treated with caffeine. After controlling for gestational age and birth weight, they had significantly lower carbon dioxide sensitivity at their initial study compared with those who did not require treatment.ConclusionCaffeine administration was associated with an increase in the ventilatory response to hypercarbia. An initial weaker ventilatory response to hypercarbia was associated with the subsequent development of apnea requiring treatment with caffeine.

Leptin Stabilizes Breathing but does not affect CO2 Chemosensitivity in Newborn Rat Pups

Premature infants have periodic breathing (PB) and chronic intermittent hypoxia (CIH) that increases during the 2nd week of life. Leptin (LEP) is a hormone produced by adipocytes and increases central CO2 chemoreceptors sensitivity. LEP is also produced by the placenta, and in newborn infants LEP levels are highest at birth and then rapidly fall. We hypothesis that the fall in LEP after birth is associated with the increase frequency of PB and apnea leading to CIH during the 2nd week of life in premature infants.ObjectiveTo determine the effect of exogenous LEP on breathing stability and CO2 sensitivity during the first week of postnatal life, using whole body flow‐through plethysmograph in freely behaving newborn rats.MethodsSprague Dawley (SD) rat pups remained with the nursing dam until the time of the experiment. On postnatal day (PD) 7, rat pups were injected intraperitoneally with either vehicle (saline/n=5), LEP6 (6 μg/gr/n=4) and LEP9 (9 μg/gr/n=5). After injection the pup was returned to nursing dam and litter mates for 30 mins followed by 30 mins in the plethysmograph prior to recording. Respiratory tracings were captured using PowerLab 8 data acquisition system (ADInstruments) and analyzed using LabChart. Ventilatory parameters; frequency, VT and Ve were compared during 21% O2, 40% O2/0% CO2, 40%O2/5%CO2 and return to 40%O2/0%CO2, and the coefficient of variation (CV) for Ve was calculated for each condition in the 3 treatment groups (SAL, LEP6, LEP9).ResultsSimilar to Stunden et al. in rat pups at PD 7–8 (Stunden et al,, Respir Physiol, 2001), frequency, VT and Ve did not significantly changed in response to hypercapnia (P=0.511), and LEP6 or LEP9 did not alter this response. However, LEP exposure did stabilize breathing with a reduction in the CV during 21%O2, 40%O2/0%CO2 and 40%O2/5%CO2 (Figure 1).ConclusionOur preliminary findings are consistent with that of Stunden et al, who reported no change in Ve in response to hypercapnia at PD 6–8 in SD rats. At this age, the hypothalamus is insensitive to the effect of leptin on feeding and metabolism (MacKay H et al., Mol Metab. 2014). Here we show that leptin does not induce nor augment CO2 chemosensitivity in newborn rats at PD7, but does at PD 18 (Gauda et al, unpublished data), and adults (Isono S et al., Anesthesiology, 2009). However our data does show that leptin stabilizes breathing during the first week of life in newborn rat pups. We speculate that unstable breathing that occurs in premature infants may be associated with the precipitous fall in leptin levels after birth.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Esketamine counters opioid-induced respiratory depression

BackgroundOpioids can produce life-threatening respiratory depression. This study tested whether subanaesthetic doses of esketamine stimulate breathing in an established human model of opioid-induced respiratory depression. MethodsIn a study with a randomised, double blind, placebo controlled, crossover design, 12 healthy, young volunteers of either sex received a dose escalating infusion of esketamine (cumulative dose 40 mg infused in 1 h) on top of remifentanil-induced respiratory depression. A population pharmacokinetic-pharmacodynamic analysis was performed with sites of drug action at baseline ventilation, ventilatory CO2-chemosensitivity, or both. ResultsRemifentanil reduced isohypercapnic ventilation (end-tidal PCO2 6.5 kPa) by approximately 40% (from 20 to 12 litre min−1) in esketamine and placebo arms of the study, through an effect on baseline ventilation and ventilatory CO2 sensitivity. The reduction in ventilation was related to a remifentanil effect on ventilatory CO2 sensitivity (~39%) and on baseline ventilation (~61%). Esketamine increased breathing through an exclusive stimulatory effect on ventilatory CO2 sensitivity. The remifentanil concentration that reduced ventilatory CO2 sensitivity by 50% (C50) was doubled at an esketamine concentration of 127 (84-191) ng ml−1 [median (interquartile range)]; the esketamine effect was rapid and driven by plasma pharmacokinetics. Placebo had no systematic effect on opioid-induced respiratory depression. ConclusionsEsketamine effectively countered remifentanil-induced respiratory depression, an effect that was attributed to an increase in remifentanil-reduced ventilatory CO2 chemosensitivity.

Respiratory Responses during Exercise in Self-contained Breathing Apparatus among Firefighters and Nonfirefighters.

BackgroundFirefighters are required to use self-contained breathing apparatus (SCBA), which impairs ventilatory mechanics. We hypothesized that firefighters have elevated arterial CO2 when using SCBA.MethodsFirefighters and controls performed a maximal exercise test on a cycle ergometer and two graded exercise tests (GXTs) at 25%, 50%, and 70% of their maximal aerobic power, once with a SCBA facemask and once with protective clothing and full SCBA.ResultsRespiratory rate increased more in controls than firefighters. Heart rate increased as a function of oxygen consumption () more in controls than firefighters. End-tidal CO2 (ETCO2) during the GXTs was not affected by work rate in either group for either condition but was higher in firefighters at all work rates in both GXTs. SCBA increased ETCO2 in controls but not firefighters.ConclusionsThe present study showed that when compared to controls, firefighters’ hypoventilate during a maximal test and GXT. The hypoventilation resulted in increased ETCO2, and presumably increased arterial CO2, during exertion. It is proposed that firefighters have altered CO2 sensitivity due to voluntary hypoventilation during training and work. Confirmation of low CO2 sensitivity and the consequence of this on performance and long-term health remain to be determined.

Conserved DNA Methylation Signatures in Early Maternal Separation and in Twins Discordant for CO2 Sensitivity

Respiratory and emotional responses to blood-acidifying inhalation of CO2 are markers of some human anxiety disorders, and can be enhanced by repeatedly cross-fostering (RCF) mouse pups from their biological mother to unrelated lactating females. Yet, these dynamics remain poorly understood. We show RCF-associated intergenerational transmission of CO2 sensitivity in normally-reared mice descending from RCF-exposed females, and describe the accompanying alterations in brain DNA methylation patterns. These epigenetic signatures were compared to DNA methylation profiles of monozygotic twins discordant for emotional reactivity to a CO2 challenge. Altered methylation was consistently associated with repeated elements and transcriptional regulatory regions among RCF-exposed animals, their normally-reared offspring, and humans with CO2 hypersensitivity. In both species, regions bearing differential methylation were associated with neurodevelopment, circulation, and response to pH acidification processes, and notably included the ASIC2 gene. Our data show that CO2 hypersensitivity is associated with specific methylation clusters and genes that subserve chemoreception and anxiety. The methylation status of genes implicated in acid-sensing functions can inform etiological and therapeutic research in this field.

Reappraising Preclinical Models of Separation Anxiety Disorder, Panic Disorder, and CO2 Sensitivity: Implications for Methodology and Translation into New Treatments.

Separation anxiety applies to multiple forms of distress responses seen in mammals during postnatal development, including separation from a caregiver. Childhood separation anxiety disorder is an important risk factor for developing panic disorder in early adulthood, and both conditions display an increased sensitivity to elevated CO2 concentrations inhaled from the air. By interfacing epidemiological, genetic, and physiological knowledge with preclinical animal research models, it is possible to decipher the mechanisms that are central to separation anxiety and panic disorders while also suggesting possible therapies. Preclinical research models allow for environmentally controlled studies of early interferences with parental care. These models have shown that different forms of early maternal separation in mice and rats induce elevated CO2 respiratory sensitivity, an important biomarker of separation anxiety and panic disorders. In mice, this is likely due to gene-environment interactions that affect multiple behavioural and physical phenotypes after exposure to this early adversity. Although several questions regarding the causal mechanism of separation anxiety and panic disorder remain unanswered, the identification and improved understanding of biomarkers that link these mental health conditions under the guise of preclinical research models in conjunction with human longitudinal cohort studies can help resolve these issues.

The role of driving factors in historical and projected carbon dynamics of upland ecosystems in Alaska.

It is important to understand how upland ecosystems of Alaska, which are estimated to occupy 84% of the state (i.e., 1,237,774km2 ), are influencing and will influence state-wide carbon (C) dynamics in the face of ongoing climate change. We coupled fire disturbance and biogeochemical models to assess the relative effects of changing atmospheric carbon dioxide (CO2 ), climate, logging and fire regimes on the historical and future C balance of upland ecosystems for the four main Landscape Conservation Cooperatives (LCCs) of Alaska. At the end of the historical period (1950-2009) of our analysis, we estimate that upland ecosystems of Alaska store ~50 Pg C (with ~90% of the C in soils), and gained 3.26 Tg C/yr. Three of the LCCs had gains in total ecosystem C storage, while the Northwest Boreal LCC lost C (-6.01Tg C/yr) because of increases in fire activity. Carbon exports from logging affected only the North Pacific LCC and represented less than 1% of the state's net primary production (NPP). The analysis for the future time period (2010-2099) consisted of six simulations driven by climate outputs from two climate models for three emission scenarios. Across the climate scenarios, total ecosystem C storage increased between 19.5 and 66.3 Tg C/yr, which represents 3.4% to 11.7% increase in Alaska upland's storage. We conducted additional simulations to attribute these responses to environmental changes. This analysis showed that atmospheric CO2 fertilization was the main driver of ecosystem C balance. By comparing future simulations with constant and with increasing atmospheric CO2 , we estimated that the sensitivity of NPP was 4.8% per 100 ppmv, but NPP becomes less sensitive to CO2 increase throughout the 21st century. Overall, our analyses suggest that the decreasing CO2 sensitivity of NPP and the increasing sensitivity of heterotrophic respiration to air temperature, in addition to the increase in C loss from wildfires weakens the C sink from upland ecosystems of Alaska and will ultimately lead to a source of CO2 to the atmosphere beyond 2100. Therefore, we conclude thatthe increasing regional C sink we estimate for the 21st century will most likely be transitional.

Potential for maternal effects on offspring CO2 sensitivities in the Atlantic silverside (Menidia menidia)

For marine fish, the influence of maternal provisioning on offspring sensitivity to high carbon dioxide (CO2) conditions remains unknown. We separately reared offspring obtained from five wild-caught Atlantic silverside (Menidia menidia) females from fertilization to 16 days post hatch under contrasting CO2 conditions (ambient: ~400μatm, acidified: ~2,300μatm), testing whether average survival during the embryo and larval stage, hatch length, final length, and growth rates were affected by CO2, female identity, or their interaction. Average trait responses did not significantly differ between treatments (CO2 or female identity), however, significant CO2×female identity interactions indicated that females produced offspring with different average CO2 sensitivities. We then examined whether differential egg provisioning with fatty acids (FA) may partially explain the observed differences in offspring CO2 sensitivities. Concentrations of 27 FAs in the unfertilized eggs of each female were measured. Cumulative absolute FA levels were negatively related to hatch length and to the log-transformed CO2 response ratio of hatch length. Eggs with lower concentrations of 20:1n9 and 22:5n3 resulted in offspring where embryo survival was negatively impacted by high CO2. Eggs with higher concentrations of 18:3n3, 18:4n3, and 22:6n3 produced shorter offspring at hatching under high CO2 conditions. These results indicate that maternal provisioning might be an additional determinant of CO2 sensitivity in fish early life stages. Acidification experiments should therefore utilize large numbers of parents from different natural conditions and, where possible, track heritage.

Reviews and syntheses: Field data to benchmark the carbon cycle models for tropical forests

Abstract. For more accurate projections of both the global carbon (C) cycle and the changing climate, a critical current need is to improve the representation of tropical forests in Earth system models. Tropical forests exchange more C, energy, and water with the atmosphere than any other class of land ecosystems. Further, tropical-forest C cycling is likely responding to the rapid global warming, intensifying water stress, and increasing atmospheric CO2 levels. Projections of the future C balance of the tropics vary widely among global models. A current effort of the modeling community, the ILAMB (International Land Model Benchmarking) project, is to compile robust observations that can be used to improve the accuracy and realism of the land models for all major biomes. Our goal with this paper is to identify field observations of tropical-forest ecosystem C stocks and fluxes, and of their long-term trends and climatic and CO2 sensitivities, that can serve this effort. We propose criteria for reference-level field data from this biome and present a set of documented examples from old-growth lowland tropical forests. We offer these as a starting point towards the goal of a regularly updated consensus set of benchmark field observations of C cycling in tropical forests.