A Rare Case of a Giant Choledochal Cyst in a Caribbean Infant.

Choledochal cysts are uncommon dilatations of the biliary tree. Giant choledochal cysts are those that exceed a maximum diameter of 10cm. Our case describes a female infant who presented to our paediatric surgery department with a three-day history of vomiting, abdominal distention, pale stool, and irritability. On palpation, she was found to have a large abdominal mass and the computed tomography (CT) scan showed a giant choledochal cyst. The patient underwent laparotomy with cholecystectomy, choledochal cyst drainage and complete excision, with hepaticojejunosotomy. At the last follow-up three years post-surgery, all growth parameters and liver enzymes were within normal ranges. To the best of our knowledge, this is the first documented case of a giant choledochal cyst in the paediatric Caribbean population.

Neuromechanical characterization of the abductor hallucis and its potential role in upright postural control.

There is growing evidence to support a role for the abductor hallucis (AH) in standing balance control; however, functional properties of the muscle that may provide more insight into AH's specific contribution to upright posture have yet to be characterized. This study was conducted to quantify functional neuromechanical properties of the AH and correlate the measures with standing balance variables. We quantified strength and voluntary activation during maximal voluntary isometric contractions of the great toe abductor in nine (3 females and 6 males) healthy, young participants. During electrically evoked twitch and tetanic contractions, we measured great toe abduction peak force and constructed a force-frequency curve. We also evaluated peak abduction force, contraction time (CT), half-relaxation time (HRT), rate of force development (RFD), and relaxation rate (RR) from twitch contractions evoked using doublet stimuli. Strength, VA, CT, HRT, RFD, and RR were correlated to centre of pressure standard deviation (COP SD) and velocity (COP VEL) variables of the traditional COP trace and its rambling and trembling components during single-legged stance. AH twitch properties (e.g., CT: 169.8 ± 32.3 ms; HRT: 124.1 ± 29.2 ms) and force-frequency curve were similar to other slow contractile muscles. Contractile speed related negatively with COP VEL, suggesting AH may be appropriate for slow, prolonged tasks such as ongoing postural balance control. Correlation coefficient outcomes for all variables were similar between rambling and trembling components. Our results provide further evidence for the importance of AH neuromechanical function for standing balance control, at least during a challenging single-legged posture.

Gata3 is required in late proneurosensory development for proper sensory cell formation and organization.

It has previously been shown that the zinc-finger transcription factor Gata3 has dynamic expression within the inner ear throughout embryonic development and is essential for cochlear neurosensory development. However, the temporal window for which Gata3 is required for proper formation of the cochlear neurosensory epithelia remains unclear. To investigate the role of Gata3 in cochlear neurosensory development in the late prosensory stages, we used the Sox2-creERT2 mouse line to target and conditionally delete Gata3 at E11.5, a timepoint before cells have fully committed to a neurosensory fate. While the inner ears of Sox2-creERT2: Gata3 f/f mice appear normal with no gross structural defects, the sensory cells in the organ of Corti are partially lost and disorganized in an increasing severity from base to apex. Additionally, spiral ganglion neurons display aberrant peripheral projections, including increased distances between radial bundles and disorganization upon reaching the organ of Corti. Furthermore, heterozygous Sox2-creERT2: Gata3 f/+ mice show a reduced aberrant phenotype in comparison to the homozygous mutant, supporting the hypothesis that Gata3 is not only required for proper formation at the later proneurosensory stage, but also that a specific expression level of Gata3 is required. Therefore, this study provides evidence that Gata3 plays a time-sensitive and dose-dependent role in the development of sensory and neuronal cells in late proneurosensory stages.

Gata3 is Required in Late Proneurosensory Development for Proper Sensory Cell Formation and Organization.

It has been previously shown that zinc-finger transcription factor Gata3 has dynamic expression within the inner ear throughout embryonic development and is essential for cochlear neurosensory development. However, the temporal window to which Gata3 is required for the formation of the cochlear neurosensory epithelia remains unclear. To investigate the role of Gata3 on cochlear neurosensory development in the late prosensory stages, we used the Sox2-cre ERT2 mouse line to target and conditionally delete Gata3 at E11.5 before the cells have fully committed to a neurosensory fate. While the inner ears of Sox2-cre ERT2 : Gata3 f/f mice appear morphologically normal, the sensory cells in the organ of Corti are partially lost and disorganized in a basal to apical gradient with the apex demonstrating the more severe phenotype. Additionally, spiral ganglion neurons display aberrant peripheral projections, such as increased distances between radial bundles and disorganization upon reaching the organ of Corti. Furthermore, heterozygous Sox2-cre ERT2 : Gata3 f/+ mice show a reduced phenotype in comparison to the homozygous mutant, supporting the concept that Gata3 is not only required for proper formation at the later proneurosensory stage, but also that a specific level of Gata3 is required. Therefore, our studies confirm that Gata3 plays a time-sensitive and dose-dependent role in the development of sensory cells in the late proneurosensory stages.

Impact of Fluid Flow Shear Stress on Osteoblast Differentiation and Cross-Talk with Articular Chondrocytes.

Bone cells, in particular osteoblasts, are capable of communication with each other during bone growth and homeostasis. More recently it has become clear that they also communicate with other cell-types; including chondrocytes in articular cartilage. One way that this process is facilitated is by interstitial fluid movement within the pericellular and extracellular matrices. This stimulus is also an important mechanical signal in skeletal tissues, and is known to generate shear stresses at the micron-scale (known as fluid flow shear stresses (FFSS)). The primary aim of this study was to develop and characterize an in vitro bone-cartilage crosstalk system, to examine the effect of FFSS on these cell types. Specifically, we evaluated the response of osteoblasts and chondrocytes to FFSS and the effect of FFSS-induced soluble factors from the former, on the latter. This system will ultimately be used to help us understand the role of subchondral bone damage in articular cartilage degeneration. We also carried out a comparison of responses between cell lines and primary murine cells in this work. Our findings demonstrate that primary cells produce a more reliable and reproducible response to FFSS. Furthermore we found that at lower magnitudes , direct FFSS produces anabolic responses in both chondrocytes and osteoblasts, whereas higher levels produce more catabolic responses. Finally we show that exposure to osteoblast-derived factors in conditioned media experiments produced similarly catabolic changes in primary chondrocytes.

Experimental and observational evidence of negative conspecific density dependence in temperate ectomycorrhizal trees.

Conspecific negative density dependence (CNDD) promotes tree species diversity by reducing recruitment near conspecific adults due to biotic feedbacks from herbivores, pathogens, or competitors. While this process is well-described in tropical forests, tests of temperate tree species range from strong positive to strong negative density dependence. To explain this, several studies have suggested that tree species traits may help predict the strength and direction of density dependence: for example, ectomycorrhizal-associated tree species typically exhibit either positive or weaker negative conspecific density dependence. More generally, the strength of density dependence may be predictably related to other species-specific ecological attributes such as shade tolerance, or the relative local abundance of a species. To test the strength of density dependence and whether it affects seedling community diversity in a temperate forest, we tracked the survival of seedlings of three ectomycorrhizal-associated species experimentally planted beneath conspecific and heterospecific adults on the Prospect Hill tract of the Harvard Forest, in Massachusetts, USA. Experimental seedling survival was always lower under conspecific adults, which increased seedling community diversity in one of six treatments. We compared these results to evidence of CNDD from observed sapling survival patterns of 28 species over approximately 8 years in an adjacent 35-ha forest plot. We tested whether species-specific estimates of CNDD were associated with mycorrhizal association, shade tolerance, and local abundance. We found evidence of significant, negative conspecific density dependence (CNDD) in 23 of 28 species, and positive conspecific density dependence in two species. Contrary to our expectations, ectomycorrhizal-associated species generally exhibited stronger (e.g., more negative) CNDD than arbuscular mycorrhizal-associated species. CNDD was also stronger in more shade-tolerant species but was not associated with local abundance. Conspecific adult trees often have a negative influence on seedling survival in temperate forests, particularly for tree species with certain traits. Here we found strong experimental and observational evidence that ectomycorrhizal-associating species consistently exhibit CNDD. Moreover, similarities in the relative strength of density dependence from experiments and observations of sapling mortality suggest a mechanistic link between negative effects of conspecific adults on seedling and sapling survival and local tree species distributions.

A comprehensive finite element model for studying Cochlear-Vestibular interaction.

We present a 3-D finite element (FE) model of the chinchilla's inner ear consisting of the entire cochlea structure and the vestibular system. The reaction of the basilar membrane to the head rotation and the reaction of ampulla to the stapes movement were investigated. These results demonstrate the existence of hearing-vestibular system interaction. They provide an explanation to the clinical finding on the coexistence between hearing loss and equilibration dysfunction. It is a preliminary, yet critical step toward the development of a comprehensive FE model of an entire ear for mechano-acoustic analysis.

The development of the O2-sensing system in an amphibious fish: consequences of variation in environmental O2 levels.

Proper development of the O2-sensing system is essential for survival. Here, we characterized the development of the O2-sensing system in the mangrove rivulus (Kryptolebias marmoratus), an amphibious fish that transitions between hypoxic aquatic environments and O2-rich terrestrial environments. We found that NECs formed in the gills and skin of K. marmoratus during embryonic development and that both NEC populations are retained from the embryonic stage to adulthood. We also found that the hyperventilatory response to acute hypoxia was present in embryonic K. marmoratus, indicating that functional O2-sensing pathways are formed during embryonic development. We then exposed embryos to aquatic normoxia, aquatic hyperoxia, aquatic hypoxia, or terrestrial conditions for the first 30 days of embryonic development and tested the hypothesis that environmental O2 availability during embryonic development modulates the development of the O2-sensing system in amphibious fishes. Surprisingly, we found that O2 availability during embryonic development had little impact on the density and morphology of NECs in the gills and skin of K. marmoratus. Collectively, our results demonstrate that, unlike the only other species of fish in which NEC development has been studied to date (i.e., zebrafish), NEC development in K. marmoratus is largely unaffected by environmental O2 levels during the embryonic stage, indicating that there is interspecies variation in O2-induced plasticity in the O2-sensing system of fishes.

The validity and reliability of ultra-short-term heart rate variability parameters and the influence of physiological covariates.

Ultra-short-term (UST) heart rate variability (HRV) metrics have increasingly been proposed as surrogates for short-term HRV metrics. However, the concurrent validity, within-day reliability, and between-day reliability of UST HRV have yet to be comprehensively documented. Thirty-six adults (18 males, age: 26 ± 5 yr, BMI: 24 ± 3 kg/m2) were recruited. Measures of HRV were quantified in a quiet-stance upright orthostatic position via three-lead electrocardiogram (ADInstruments, FE232 BioAmp). All short-term data recordings were 300 s in length and five UST time points (i.e., 30 s, 60 s, 120 s, 180 s, and 240 s) were extracted from the original 300-s recording. Bland-Altman plots with 95% limits of agreement, repeated measures ANOVA and two-tailed paired t tests demarcated differences between UST and short-term recordings. Linear regressions, coefficient of variation, intraclass correlation coefficients, and other tests examined the validity and reliability in both time- and frequency domains. No group differences were noted between all short-term and UST measures, for either time- (all P > 0.202) or frequency-domain metrics (all P > 0.086). A longer recording duration was associated with augmented validity and reliability, which was less impacted by confounding influences from physiological variables (e.g., respiration rate, carbon dioxide end-tidals, and blood pressure). Conclusively, heart rate, time-domain, and relative frequency-domain HRV metrics were acceptable with recordings greater or equal to 60 s, 240 s, and 300 s, respectively. Future studies employing UST HRV metrics should thoroughly understand the methodological requirements to obtain accurate results. Moreover, a conservative approach should be utilized regarding the minimum acceptable recording duration, which ensures valid/reliable HRV estimates are obtained.NEW & NOTEWORTHY A one size fits all methodological approach to quantify HRV metrics appears to be inappropriate, where study design considerations need to be conducted upon a variable-by-variable basis. The present results found 60 s (heart rate), 240 s (time-domain parameters), and 300 s (relative frequency-domain parameters) were required to obtain accurate and reproducible metrics. The lower validity/reliability of the ultra-short-term metrics was attributable to measurement error and/or confounding from extraneous physiological influences (i.e., respiratory and hemodynamic variables).

The impact of high- and moderate-intensity exercise on near-point of convergence metrics.

Background: Near point of convergence (NPC) assesses the vergence ability of the visuo-oculomotor system; however, little is known regarding: the extent and duration exercise impacts NPC and the between- and within-day reliability of NPC metrics.Methods: An accommodative ruler with a miniature Snellen chart was placed upon the philtrum (upper lip). Participants (n=9) focused upon a 'V' sized 20/20, while the chart was moved at ~1-2 cm/s toward and away from the eyes (twice in each direction). Testing commenced at 8:00am with NPC measures being collected at baseline before three randomized conditions with serial follow-ups occurring at six post-condition timepoints (0-8 hours following). The conditions consisted of 25-minutes high-intensity intervals (10, one-minute intervals at ~85-90% heart-rate reserve), 45-minutes of moderate-intensity exercise (at ~50-60% heart-rate reserve), and a control condition (30-minutes quiet rest).Results: NPC was not impacted across any of the three conditions (all p > .59). Additionally, NPC measures between baseline conditions and across the control condition displayed very high levels of within-day and between-day reliability (coefficient of variation <3.8%).Conclusions: Future NPC measures using an accommodative ruler can be taken immediately following exercise and may be pertinent as a complementary tool in the future sideline screening of concussion.

Temporal evolution of neurovascular coupling recovery following moderate- and high-intensity exercise.

PURPOSE: Studies examining neurovascular coupling (NVC) require participants to refrain from exercise for 12-24 hours. However, there is a paucity of empirical evidence for this restriction. The objectives for this study were to delineate the time-course recovery of NVC metrics following exercise and establish the NVC within- and between-day reliability. METHODS: Nine participants completed a complex visual search paradigm to assess NVC via transcranial Doppler ultrasound of the posterior cerebral artery blood velocity (PCA). Measurements were performed prior to and throughout the 8-hour recovery period following three randomized conditions: 45 minutes of moderate-intensity exercise (at 50% heart-rate reserve), 30 minutes high-intensity intervals (10, 1-minute intervals at 85% heart-rate reserve), and control (30 minutes quiet rest). In each condition, baseline measures were collected at 8:00am with serial follow-ups at hours zero, one, two, four, six, and eight. RESULTS: Area-under-the-curve and time-to-peak PCA velocity during the visual search were attenuated at hour zero following high-intensity intervals (all p < 0.05); however, these NVC metrics recovered at hour one (all p > 0.13). Conversely, baseline PCA velocity, peak PCA velocity, and the relative percent increase were not different following high-intensity intervals compared to baseline (all p > 0.26). No NVC metrics differed from baseline following both moderate exercise and control conditions (all p > 0.24). The majority of the NVC parameters demonstrated high levels of reliability (intraclass correlation coefficient: >0.90). CONCLUSION: Future NVC assessments can take place a minimum of one hour following exercise. Moreover, all metrics did not change across the control condition, therefore future studies using this methodology can reliably quantify NVC between 8:00am and 7:00 pm.

Fzd3 Expression Within Inner Ear Afferent Neurons Is Necessary for Central Pathfinding.

During development the afferent neurons of the inner ear make precise wiring decisions in the hindbrain reflective of their topographic distribution in the periphery. This is critical for the formation of sensory maps capable of faithfully processing both auditory and vestibular input. Disorganized central projections of inner ear afferents in Fzd3 null mice indicate Wnt/PCP signaling is involved in this process and ear transplantation in Xenopus indicates that Fzd3 is necessary in the ear but not the hindbrain for proper afferent navigation. However, it remains unclear in which cell type of the inner ear Fzd3 expression is influencing the guidance of inner ear afferents to their proper synaptic targets in the hindbrain. We utilized Atoh1-cre and Neurod1-cre mouse lines to conditionally knockout Fzd3 within the mechanosensory hair cells of the organ of Corti and within the inner ear afferents, respectively. Following conditional deletion of Fzd3 within the hair cells, the central topographic distribution of inner ear afferents was maintained with no gross morphological defects. In contrast, conditional deletion of Fzd3 within inner ear afferents leads to central pathfinding defects of both cochlear and vestibular afferents. Here, we show that Fzd3 is acting in a cell autonomous manner within inner ear afferents to regulate central pathfinding within the hindbrain.

Fluctuating environments during early development can limit adult phenotypic flexibility: insights from an amphibious fish.

The interaction between developmental plasticity and the capacity for reversible acclimation (phenotypic flexibility) is poorly understood, particularly in organisms exposed to fluctuating environments. We used an amphibious killifish (Kryptolebias marmoratus) to test the hypotheses that organisms reared in fluctuating environments (i) will make no developmental changes to suit any one environment because fixing traits to suit one environment could be maladaptive for another, and (ii) will be highly phenotypically flexible as adults because their early life experiences predict high environmental variability in the future. We reared fish under constant (water) or fluctuating (water-air) environments until adulthood and assessed a suite of traits along the oxygen cascade (e.g. neuroepithelial cell density and size, cutaneous capillarity, gill morphology, ventricle size, red muscle morphometrics, terrestrial locomotor performance). To evaluate the capacity for phenotypic flexibility, a subset of adult fish from each rearing condition was then air-exposed for 14 days before the same traits were measured. In support of the developmental plasticity hypothesis, traits involved with O2 sensing and uptake were largely unaffected by water-air fluctuations during early life, but we found marked developmental changes in traits related to O2 transport, utilization and locomotor performance. In contrast, we found no evidence supporting the phenotypic flexibility hypothesis. Adult fish from both rearing conditions exhibited the same degree of phenotypic flexibility in various O2 sensing- and uptake-related traits. In other cases, water-air fluctuations attenuated adult phenotypic flexibility despite the fact that phenotypic flexibility is hypothesized to be favoured when environments fluctuate. Overall, we conclude that exposure to environmental fluctuations during development in K. marmoratus can dramatically alter the constitutive adult phenotype, as well as diminish the scope for phenotypic flexibility in later life.

Effects of high-intensity intervals and moderate-intensity exercise on baroreceptor sensitivity and heart rate variability during recovery.

Numerous studies have examined heart rate variability (HRV) and cardiac baroreceptor sensitivity (BRS) variables during recovery both acutely (under 3 h) and long-term (24, 48, and 72 h) postexercise. However, there is little literature examining HRV and BRS measures between these timepoints. Spontaneous short-term HRV and cardiac BRS measures were collected in 9 participants before and at zero, 1, 2, 4, 6, and 8 h after 3 separate conditions: moderate-intensity continuous exercise (MICE; 45 min at 50% heart rate reserve), high-intensity interval exercise (HIIE; 25 min including ten 1-min intervals at 85% heart rate reserve), and control (30 min quiet rest). HRV measures in the time domain were only affected immediately following HIIE and MICE at hour zero (all p < 0.043), whereas frequency-domain metrics were unaltered (all p > 0.102). These measures were highly consistent across the control day (all p > 0.420). Cardiac BRS was assessed via low-frequency (LF) gain, and revealed reductions following HIIE at hour zero (p < 0.012). Cardiac BRS LF gain remained consistent following MICE and control interventions (all p > 0.280). The common practice of waiting 12 to 24 h is overly conservative as the current findings demonstrate measures return to baseline at ∼60 min after exercise. Moreover, these metrics demonstrated high levels of within- and between-day reliability. Novelty Previously a 12-h minimum restriction from exercise was required before participation in HRV/BRS studies. Recovery from moderate-intensity exercise for HRV and BRS metrics was <60 min; whereas, high-intensity intervals led to alterations for approximately 60 min. Spontaneous HRV and cardiac BRS demonstrated high levels of within-day reproducibility.

Modulation of vestibular-evoked responses prior to simple and complex arm movements.

During destabilizing, voluntary arm movements, the vestibular system provides sensory cues related to head motion that are necessary to preserve upright balance. Although sensorimotor processing increases in accordance with task complexity during the preparation phase of reaching, it is unclear whether vestibular signals are also enhanced when maintaining postural control prior to the execution of a voluntary movement. To probe whether vestibular cues are a component of complexity-related increases in sensorimotor processing during movement preparation, vestibular-evoked responses to stochastic (0-25 Hz; root mean square = 1 mA) binaural, bipolar electrical vestibular stimulation (EVS) were examined. These responses were assessed using cumulant density function estimates in the upper and lower limbs prior to ballistic arm movements of varying complexity in both standing (experiment 1) and seated (experiment 2) conditions. In experiment 1, EVS-electromyography (EMG) cumulant density estimates surpassed 95% confidence intervals for biceps and triceps brachii, as well as the left and right medial gastrocnemius. For the latter two muscles, the responses were enhanced 10-18% with increased movement complexity. In experiment 2, the EVS-EMG cumulant density estimates also surpassed 95% confidence intervals in the upper limb, confirming the presence of vestibular-evoked responses while seated; however, the amplitude was significantly less than standing. This study demonstrates the vestibular system contributes to postural stability during the preparation phase of reaching. As such, vestibular-driven signals may be used to update an internal model for upcoming reaching tasks or to prepare for imminent postural disturbances.

Hot weather increases competition between dairy cows at the drinker.

Heat-stressed dairy cows on pasture will compete for resources that aid cooling, but it is not known how heat stress affects the competition for water by indoor-housed cows. The aim of this observational study was to evaluate how heat stress affects the behavior of indoor-housed cows at the drinker at both group and cow levels. For 3 wk after calving, cows were housed in a dynamic group of 20 animals in a pen with 12 electronic feed bins, 2 electronic water bins, and 24 freestalls. A total of 69 lactating Holstein dairy cows were enrolled over the 59-d study. The electronic water bins recorded time spent at the drinker, frequency of visits, water intake, and competitive events for 24 h/d. Competitive events were quantified using the number of replacements (recorded when there was a ≤29-s interval between 2 cows sequentially visiting the same drinker). The number of replacements a cow was involved in was used to determine her level of competitive success at the drinker (low, medium, high). The temperature-humidity index (THI) was recorded by the local weather station, and moving averages for daily maximum THI over a 3-d period were calculated. For the analysis of time spent at the drinker, frequency of visits, and water intake, the measures from all cows were averaged to create 1 observation per day, and the number of replacements at the drinker was summed. A linear regression was performed to determine the relationship between THI and group-level drinking behavior. At the cow level, a repeated measures mixed model, with fixed effects of level of competitive success, milk yield, and 3-d maximum THI and a first-order autoregressive covariance structure, was used to determine how increasing THI affects the drinking behavior of individual cows based on their level of competitive success. Feed intake was included as a fixed effect in the water intake model. We found that, with increasing THI, cows drank more water, spent more time at the drinker, made more visits to the drinker, and engaged in more competitive events at the drinker. In exploratory analysis, we found that cows with low competitive success at the drinker shifted their drinking behavior to avoid the drinker at the hottest and most competitive time of day. These results indicate that behavior can be used to indicate when cows feel hot. These measures may be of practical value in deciding when to provide cooling, especially for farms where attendance at the drinker can be monitored electronically.

Ageing impacts phenotypic flexibility in an air-acclimated amphibious fish.

The ability to tolerate environmental change may decline as fishes age. We tested the hypothesis that ageing influences the scope for phenotypic flexibility in the mangrove rivulus (Kryptolebias marmoratus), an amphibious fish that transitions between two vastly different environments, water and land. We found that older fish (4-6 years old) exhibited marked signs of ageing; older fish were reproductively senescent, had reduced fin regenerative capacity and body condition, and exhibited atrophy of both oxidative and glycolytic muscle fibers relative to younger adult fish (1-2 years old). However, age did not affect routine O2 consumption. We then acclimated adult fish (1-6 years) to water (control) or air for 10 days to assess the scope for phenotypic flexibility in response to terrestrial exposure. In support of our hypothesis, we found that older air-acclimated fish had a diminished scope for gill remodeling relative to younger fish. We also found that older fish exhibited poorer terrestrial locomotor performance relative to younger adult fish, particularly when acclimated to air. Our results indicate that ageing diminishes skeletal muscle integrity and locomotor performance of amphibious fishes, and may, therefore, impair terrestrial foraging ability, predator avoidance, or dispersal across the terrestrial environment. Remarkably, older fish voluntarily left water to a similar degree as younger fish despite the age-related deterioration of traits important for terrestrial life.

Hydrogen sulphide toxicity and the importance of amphibious behaviour in a mangrove fish inhabiting sulphide-rich habitats.

We investigated amphibious behaviour, hydrogen sulphide (H2S) tolerance, and the mechanism of H2S toxicity in the amphibious mangrove rivulus (Kryptolebias marmoratus). We found that fish emersed (left water) in response to acutely elevated [H2S] (~ 130-200 µmol l-1). The emersion response to H2S may be influenced by prior acclimation history due to acclimation-induced alterations in gill morphology and/or the density and size of neuroepithelial cells (NECs) on the gills and skin. Thus, we acclimated fish to water (control), H2S-rich water, or air and tested the hypotheses that acclimation history influences H2S sensitivity due to acclimation-induced changes in (i) gill surface area and/or (ii) NEC density and/or size. Air-acclimated fish emersed at significantly lower [H2S] relative to fish acclimated to control or H2S-rich water, but exhibited no change in gill surface area or in NEC density or size in the gills or skin. Despite possessing exceptional H2S tolerance, all fish lost equilibrium when unable to emerse from environments containing extremely elevated [H2S] (2272 ± 46 µmol l-1). Consequently, we tested the hypothesis that impaired blood oxygen transport (i.e., sulphemoglobin formation) causes H2S toxicity in amphibious fishes. In vitro exposure of red blood cells to physiologically relevant [H2S] did not cause a substantial increase in sulphemoglobin formation. We found evidence, however, for an alternative hypothesis that H2S toxicity is caused by impaired oxidative phosphorylation (i.e., cytochrome c oxidase inhibition). Collectively, our results show that amphibious behaviour is critical for the survival of K. marmoratus in H2S-rich environments as fish experience impaired oxidative phosphorylation when unable to emerse.

Technical note: Using an electronic drinker to monitor competition in dairy cows.

The objective of this study was to determine whether data from an electronic drinking system that tracks individual animal attendance with ear tag transponders could be used to detect social competition between dairy cows at the drinker. Specifically, we sought to identify the interval between one cow leaving the drinker and another cow taking her place that most accurately identified competitive replacements (when physical contact initiated by one cow causes the other to entirely remove her head from the drinker and the initiator subsequently places her head in the same drinker). The optimal interval to accurately identify replacements at the drinker was determined using a receiver operating characteristic curve and by evaluating the sensitivity (Se), specificity (Sp), and confusion matrix values (true positives, true negatives, false positives, false negatives) of different thresholds identified by 3 metrics: the Youden index, the point closest to (0,1) on the curve, and the point where Se roughly equals Sp. Lactating Holstein dairy cows (n = 20) were monitored for 4 consecutive 24-h periods by video recording and with an electronic drinking system. Two periods were used to identify the optimal interval (the baseline set), and the other 2 periods were used to validate the interval (the validation set). The occurrence of a replacement identified by video was paired with the interval between drinking events of 2 cows at the same electronic drinker to identify the interval that best predicted replacement events. Based on the low prevalence of replacement events compared with nonreplacement events, the interval with the lowest number of false positives was considered optimal. Of the 3 potential metrics, the point on the receiver operating characteristic curve where Se and Sp were roughly equal yielded the fewest false positives. The optimal interval to identify replacements at the drinker was ≤29 s, with 82% Se and 83% Sp in the baseline set. This interval was found to have 85% Se and 89% Sp when applied to the validation set. These results indicate that social competition between dairy cows at the drinker can be accurately measured with an electronic drinking system.

Inhibition of upper small intestinal mTOR lowers plasma glucose levels by inhibiting glucose production.

Glucose homeostasis is partly controlled by the energy sensor mechanistic target of rapamycin (mTOR) in the muscle and liver. However, whether mTOR in the small intestine affects glucose homeostasis in vivo remains unknown. Here, we first report that delivery of rapamycin or an adenovirus encoding the dominant negative acting mTOR-mutated protein into the upper small intestine is sufficient to inhibit small intestinal mTOR signaling and lower glucose production in rodents with high fat diet-induced insulin resistance. Second, we found that molecular activation of small intestinal mTOR blunts the glucose-lowering effect of the oral anti-diabetic agent metformin, while inhibiting small intestinal mTOR alone lowers plasma glucose levels by inhibiting glucose production in rodents with diabetes as well. Thus, these findings illustrate that inhibiting upper small intestinal mTOR is sufficient and necessary to lower glucose production and enhance glucose homeostasis, and thereby unveil a previously unappreciated glucose-lowering effect of small intestinal mTOR.