Does intensity matter? A randomized crossover study of the role of acute exercise intensity on cognitive performance and motor speed and accuracy.

There is a well-recognized, yet nuanced, positive relationship between acute physical activity and cognitive function. However, the precise impact of exercise intensity remains ambiguous. We tested learning and memory, working memory and processing speed, and motor speed and accuracy across three distinct exercise intensities. A sample of 207 participants (100 female) between 18 and 44 years (mean age: 22.5±3.7years) completed all study procedures. Utilizing a within-subjects, cross-over design, participants completed moderate (35% VO2 Max), vigorous (70% VO2 Max), and sedentary (no exercise) conditions. Cognitive and motor assessments, including the Paced Auditory Serial Addition Test (PASAT), Rey Auditory Verbal Learning Test (RAVLT), Typing Speed Test, and Ten Key Data Entry Task, were conducted approximately 60min post-exercise. There were no significant differences in primary cognitive or motor outcome measures across the three exercise intensities, even with the study being strongly powered. There was, however, a small difference on the fastest trial of the PASAT, where vigorous-intensity exercise yielded slightly better performance compared to both sedentary and moderate-intensity exercise. This effect was no longer significant when including VO2 Max or maximum heart rate as indicators of fitness. There were no interactions on outcome variables by exercise intensity when including biological sex in the models. Thus, a single bout of acute exercise, regardless of its intensity, did not alter cognitive and motor performance when measured approximately 1h post-exercise. Findings highlight the importance of large samples and suggest that the temporal dynamics post-exercise might play a pivotal role in cognitive outcomes.

Opto-SICM framework combines optogenetics with scanning ion conductance microscopy for probing cell-to-cell contacts.

We present a novel framework, Opto-SICM, for studies of cellular interactions in live cells with high spatiotemporal resolution. The approach combines scanning ion conductance microscopy, SICM, and cell-type-specific optogenetic interrogation. Light-excitable cardiac fibroblasts (FB) and myofibroblasts (myoFB) were plated together with non-modified cardiomyocytes (CM) and then paced with periodic illumination. Opto-SICM reveals the extent of FB/myoFB-CM cell-cell contacts and the dynamic changes over time not visible by optical microscopy. FB-CM pairs have lower gap junctional expression of connexin-43 and higher contact dynamism compared to myoFB-CM pairs. The responsiveness of CM to pacing via FB/myoFB depends on the dynamics of the contact but not on the area. The non-responding pairs have higher net cell-cell movement at the contact. These findings are relevant to cardiac disease states, where adverse remodeling leads to abnormal electrical excitation of CM. The Opto-SICM framework can be deployed to offer new insights on cellular and subcellular interactions in various cell types, in real-time.

Nanopore sequencing of DNA-barcoded probes for highly multiplexed detection of microRNA, proteins and small biomarkers.

There is an unmet need to develop low-cost, rapid and highly multiplexed diagnostic technology platforms for quantitatively detecting blood biomarkers to advance clinical diagnostics beyond the single biomarker model. Here we perform nanopore sequencing of DNA-barcoded molecular probes engineered to recognize a panel of analytes. This allows for highly multiplexed and simultaneous quantitative detection of at least 40 targets, such as microRNAs, proteins and neurotransmitters, on the basis of the translocation dynamics of each probe as it passes through a nanopore. Our workflow is built around a commercially available MinION sequencing device, offering a one-hour turnaround time from sample preparation to results. We also demonstrate that the strategy can directly detect cardiovascular disease-associated microRNA from human serum without extraction or amplification. Due to the modularity of barcoded probes, the number and type of targets detected can be significantly expanded.

Multiple generations of antibiotic exposure and isolation influence host fitness and the microbiome in a model zooplankton species.

Chronic antibiotic exposure impacts host health through changes to the microbiome. The detrimental effects of antibiotic perturbation on microbiome structure and function after one host generation of exposure have been well-studied, but less is understood about multigenerational effects of antibiotic exposure and subsequent recovery. In this study, we examined microbiome composition and host fitness across five generations of exposure to antibiotics in the model zooplankton host Daphnia magna. By utilizing a split-brood design where half of the offspring from antibiotic-exposed parents were allowed to recover and half were maintained in antibiotics, we examined recovery and resilience of the microbiome. Unexpectedly, we discovered that isolation of single host individuals across generations exerted a strong effect on microbiome composition, with microbiome diversity decreasing over generations regardless of treatment, while host body size and cumulative reproduction increased across generations. Though antibiotics did cause substantial changes to microbiome composition within a generation, recovery generally occurred in one generation regardless of the number of prior generations spent in antibiotics. Our results demonstrate that isolation of individual hosts leads to stochastic extinction of less abundant taxa in the microbiome, suggesting that these taxa are likely maintained via transmission in host populations.

Daily feeding rhythm linked to microbiome composition in two zooplankton species.

Host-associated microbial communities are impacted by external and within-host factors, i.e., diet and feeding behavior. For organisms known to have a circadian rhythm in feeding behavior, microbiome composition is likely impacted by the different rates of microbe introduction and removal across a daily cycle, in addition to any diet-induced changes in microbial interactions. Here, we measured feeding behavior and used 16S rRNA sequencing to compare the microbial community across a diel cycle in two distantly related species of Daphnia, that differ in their life history traits, to assess how daily feeding patterns impact microbiome composition. We find that Daphnia species reared under similar laboratory conditions have significantly different microbial communities. Additionally, we reveal that Daphnia have daily differences in their microbial composition that correspond with feeding behavior, such that there is greater microbiome diversity at night during the host's active feeding phase. These results highlight that zooplankton microbiomes are relatively distinct and are likely influenced by host phylogeny.

Targeted Manipulation of Abundant and Rare Taxa in the Daphnia magna Microbiota with Antibiotics Impacts Host Fitness Differentially.

Host-associated microbes contribute to host fitness, but it is unclear whether these contributions are from rare keystone taxa, numerically abundant taxa, or interactions among community members. Experimental perturbation of the microbiota can highlight functionally important taxa; however, this approach is primarily applied in systems with complex communities where the perturbation affects hundreds of taxa, making it difficult to pinpoint contributions of key community members. Here, we use the ecological model organism Daphnia magna to examine the importance of rare and abundant taxa by perturbing its relatively simple microbiota with targeted antibiotics. We used sublethal antibiotic doses to target either rare or abundant members across two temperatures and then measured key host life history metrics and shifts in microbial community composition. We find that removal of abundant taxa had greater impacts on host fitness than did removal of rare taxa and that the abundances of nontarget taxa were impacted by antibiotic treatment, suggesting that no rare keystone taxa exist in the Daphnia magna microbiota but that microbe-microbe interactions may play a role in host fitness. We also find that microbial community composition was impacted by antibiotics differently across temperatures, indicating that ecological context shapes within-host microbial responses and effects on host fitness.IMPORTANCE Understanding the contributions of rare and abundant taxa to host fitness is an outstanding question in host microbial ecology. In this study, we use the model zooplankton Daphnia magna and its relatively simple cohort of bacterial taxa to disentangle the roles of distinct taxa in host life history metrics, using a suite of antibiotics to selectively reduce the abundance of functionally important taxa. We also examine how environmental context shapes the importance of these bacterial taxa in host fitness.

Exosomes: From Potential Culprits to New Therapeutic Promise in the Setting of Cardiac Fibrosis.

Fibrosis is a significant global health problem associated with many inflammatory and degenerative diseases affecting multiple organs, individually or simultaneously. Fibrosis develops when extracellular matrix (ECM) remodeling becomes excessive or uncontrolled and is associated with nearly all forms of heart disease. Cardiac fibroblasts and myofibroblasts are the main effectors of ECM deposition and scar formation. The heart is a complex multicellular organ, where the various resident cell types communicate between themselves and with cells of the blood and immune systems. Exosomes, which are small extracellular vesicles, (EVs), contribute to cell-to-cell communication and their pathophysiological relevance and therapeutic potential is emerging. Here, we will critically review the role of endogenous exosomes as possible fibrosis mediators and discuss the possibility of using stem cell-derived and/or engineered exosomes as anti-fibrotic agents.

Characterization of key bacterial species in the Daphnia magna microbiota using shotgun metagenomics.

The keystone zooplankton Daphnia magna has recently been used as a model system for understanding host-microbiota interactions. However, the bacterial species present and functions associated with their genomes are not well understood. In order to understand potential functions of these species, we combined 16S rRNA sequencing and shotgun metagenomics to characterize the whole-organism microbiota of Daphnia magna. We assembled five potentially novel metagenome-assembled genomes (MAGs) of core bacteria in Daphnia magna. Genes involved in host colonization and immune system evasion were detected across the MAGs. Some metabolic pathways were specific to some MAGs, including sulfur oxidation, nitrate reduction, and flagellar assembly. Amino acid exporters were identified in MAGs identified as important for host fitness, and pathways for key vitamin biosynthesis and export were identified across MAGs. In total, our examination of functions in these MAGs shows a diversity of nutrient acquisition and metabolism pathways present that may benefit the host, as well as genomic signatures of host association and immune system evasion.

Frequency and circumstances of falls for people with Charcot-Marie-Tooth disease: A cross sectional survey.

OBJECTIVE: People with Charcot-Marie-Tooth (CMT) disease may be at risk of falls due to distal muscle weakness and sensory impairment. We aimed to understand the frequency of falls in a cohort, where they occurred, injury and to what people attributed the possible cause. DESIGN: A cross-sectional survey design was used. It was sent by post to ascertain the frequency of falls and near falls, plus the circumstances of the last three falls events they could recall. SETTING: The survey was administered to 252 people with CMT who were patients at a specialist neuromuscular centre. PARTICIPANTS: People were approached who had a clinical diagnosis by a neuromuscular specialist neurologist. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURE: The Falls Event Questionnaire. RESULTS: Responses were received from 107 people with CMT. Falls and near falls were reported by 86% of survey respondents. The majority of falls occurred at home, and muscular weakness appeared to be blamed the most, with participants describing tripping due to foot drop or joints giving way. A similar pattern was observed for near falls, but they tended to be more frequent. Moderate and major injuries were relatively rare, with the majority of falls resulting in scrapes and bruises or the faller feeling shaken by the experience. DISCUSSION: The impact of falls is far reaching, in terms of injury risk and the consequences of the fear of falling. Interventions to improve balance and reduce falls are an important rehabilitation focus for people with CMT.

Dysregulated Zn2+ homeostasis impairs cardiac type-2 ryanodine receptor and mitsugumin 23 functions, leading to sarcoplasmic reticulum Ca2+ leakage.

Aberrant Zn2+ homeostasis is associated with dysregulated intracellular Ca2+ release, resulting in chronic heart failure. In the failing heart a small population of cardiac ryanodine receptors (RyR2) displays sub-conductance-state gating leading to Ca2+ leakage from sarcoplasmic reticulum (SR) stores, which impairs cardiac contractility. Previous evidence suggests contribution of RyR2-independent Ca2+ leakage through an uncharacterized mechanism. We sought to examine the role of Zn2+ in shaping intracellular Ca2+ release in cardiac muscle. Cardiac SR vesicles prepared from sheep or mouse ventricular tissue were incorporated into phospholipid bilayers under voltage-clamp conditions, and the direct action of Zn2+ on RyR2 channel function was examined. Under diastolic conditions, the addition of pathophysiological concentrations of Zn2+ (≥2 nm) caused dysregulated RyR2-channel openings. Our data also revealed that RyR2 channels are not the only SR Ca2+-permeable channels regulated by Zn2+ Elevating the cytosolic Zn2+ concentration to 1 nm increased the activity of the transmembrane protein mitsugumin 23 (MG23). The current amplitude of the MG23 full-open state was consistent with that previously reported for RyR2 sub-conductance gating, suggesting that in heart failure in which Zn2+ levels are elevated, RyR2 channels do not gate in a sub-conductance state, but rather MG23-gating becomes more apparent. We also show that in H9C2 cells exposed to ischemic conditions, intracellular Zn2+ levels are elevated, coinciding with increased MG23 expression. In conclusion, these data suggest that dysregulated Zn2+ homeostasis alters the function of both RyR2 and MG23 and that both ion channels play a key role in diastolic SR Ca2+ leakage.

Investigation of road salts and biotic stressors on freshwater wetland communities.

The application of road deicing salts has led to the salinization of freshwater ecosystems in northern regions worldwide. Increased chloride concentrations in lakes, streams, ponds, and wetlands may negatively affect freshwater biota, potentially threatening ecosystem services. In an effort to reduce the effects of road salt, operators have increased the use of salt alternatives, yet we lack an understanding of how these deicers affect aquatic communities. We examined the direct and indirect effects of the most commonly used road salt (NaCl) and a proprietary salt mixture (NaCl, KCl, MgCl2), at three environmentally relevant concentrations (150, 470, and 780 mg Cl-/L) on freshwater wetland communities in combination with one of three biotic stressors (control, predator cues, and competitors). The communities contained periphyton, phytoplankton, zooplankton, and two tadpole species (American toads, Anaxyrus americanus; wood frogs, Lithobates sylvaticus). Overall, we found the two road salts did not interact with the natural stressors. Both salts decreased pH and reduced zooplankton abundance. The strong decrease in zooplankton abundance in the highest NaCl concentration caused a trophic cascade that resulted in increased phytoplankton abundance. The highest NaCl concentration also reduced toad activity. For the biotic stressors, predatory stress decreased whereas competitive stress increased the activity of both tadpole species. Wood frog survival, time to metamorphosis, and mass at metamorphosis all decreased under competitive stress whereas toad time to metamorphosis increased and mass at metamorphosis decreased. Road salts and biotic stressors can both affect freshwater communities, but their effects are not interactive.

Exploring the biophysical evidence that mammalian two-pore channels are NAADP-activated calcium-permeable channels.

Nicotinic acid adenine dinucleotide phosphate (NAADP) potently releases Ca(2+) from acidic intracellular endolysosomal Ca(2+) stores. It is widely accepted that two types of two-pore channels, termed TPC1 and TPC2, are responsible for the NAADP-mediated Ca(2+) release but the underlying mechanisms regulating their gating appear to be different. For example, although both TPC1 and TPC2 are activated by NAADP, TPC1 appears to be additionally regulated by cytosolic Ca(2+) . Ion conduction and permeability also differ markedly. TPC1 and TPC2 are permeable to a range of cations although biophysical experiments suggest that TPC2 is slightly more selective for Ca(2+) over K(+) than TPC1 and hence capable of releasing greater quantities of Ca(2+) from acidic stores. TPC1 is also permeable to H(+) and therefore may play a role in regulating lysosomal and cytosolic pH, possibly creating localised acidic domains. The significantly different gating and ion conducting properties of TPC1 and TPC2 suggest that these two ion channels may play complementary physiological roles as Ca(2+) -release channels of the endolysosomal system.

Minimizing errors associated with calculating the location of the helical axis for spinal motions.

One of the more common comparative tools used to quantify the motion of the vertebral joint is the orientation and position of the (finite) helical axis of motion as well as the amount of translation along, and rotation about, this axis. A survey of recent studies that utilize the helical axis of motion to compare motion before and after total disc replacement reveals a lack of concern for the relative errors associated with this metric. Indeed, intrinsic algorithmic and experimental errors that arise when interpreting motion tracking data can easily lead to a misinterpretation of the changes caused by replacement disc devices. While previous studies examining these errors exist, most have overlooked the errors associated with the determination of the location of the helical axis and its intersection with a chosen plane. The purpose of the study presented in this paper was to evaluate the sensitivity and reliability of the helical axis of motion as a comparative tool for kinematically evaluating spinal prostheses devices. To this end, we simulated a typical spine biomechanics testing experiment to investigate the accuracy of calculating the helical axis and its associated parameters using several popular algorithms. The resultant data motivated the development of a new algorithm that is a hybrid of two existing algorithms. The improved accuracy of this hybrid method made it possible to quantify some of the changes to the kinematics of a spinal unit that are induced by distinct placements of a total disc replacement.

Morbidity data collection in general practice: experience in the South Eastern Health Board 1998-1999.

A joint project between general practitioners and the South Eastern Health Board resulted in the establishment of a network of computerised practices collecting morbidity data in 1998 - 1999. Five practices established age sex registers for public and private patients. The International Classification of Primary Care 2 (ICPC-2) was used to define inclusion criteria for a range of illnesses relevant to primary care and public health. Problems arose in validating and extracting data in three practices. Disease prevalence for 17 illnesses was established for two practices only. The project clearly established the problems associated with morbidity data collection in general practice which include absence of a national patient registration system, absence of a national electronic messaging standard, difficulties extracting data from practice software systems and the need for a high level of dedicated staff and resources to implement such a project.

Facilitating effective learning during clinical placement.

The aim of this pilot study was to employ a qualitative research methodology to explore students' perceptions of learning and assessment of that learning whilst on clinical placement. The students' perceptions were sampled by questionnaire both pre- and post-placement (ten weeks block placement). The analysis of the data was undertaken using principles from Grounded Theory (Strauss and Corbin 1990). The analysis yielded a number of categories and bipolar paradigms relating to learning and assessment. However little change was seen between comments made pre and post-placement. The implications of this analysis are discussed.

A comparison of fixed and disposable head Heaf guns.

The objective of the study was to compare the newly introduced Model 2000 disposable-head Heaf gun with the currently used fixed-head gun. The study simultaneously Heaf-tested 1410 school children on both forearms using a different gun on each arm; the tests were read blind. The results were similar for both Heaf guns. The fixed-head gun gave a stronger reaction in 202 (14%) children by one Heaf grade. The disposable head gun gave a stronger reaction in 116 (8%) children, by one Heaf grade in 113 and 2 Heaf grades in 3 children. kappa = 0.607 (95% confidence interval [CI] = +/- 0.038), indicating good agreement between the tests. chi 2 = 0.177, d.f. = 1, P = 0.7 NS. Given the practical advantages of Heaf skin testing for primary screening, it is likely that practitioners will replace the fixed-head gun with one that avoids sterilisation problems. The small variation between the tests was not considered clinically important, while the advantages of the new test were considerable. It is therefore recommended that the Model 2000 Heaf gun replace the fixed-head gun for primary screening.

A geographical distribution of mortality and deprivation.

Because of the paucity of data on the geographical distribution of mortality and deprivation in the Republic of Ireland, we calculated SMRs for each county for the years 1982-6. Two measures of mortality were used, all cause and avoidable mortality. Two indexes of deprivation were also calculated for each county, Townsend's and a new Irish index. Significant differences in mortality exist between counties. Counties that had SMRs significantly below the national standard of 100 were Meath, Donegal and Galway. Counties with significantly high SMRs were Louth, Limerick and Westmeath. County SMRs were correlated with both deprivation indexes and no significant correlation was found to explain the apparent differences in mortality.

The Kilkenny Post-Primary Schools Survey--a survey of knowledge, attitudes and behaviour relevant to non-communicable diseases.

This survey was carried out in the Spring of 1987 on a stratified random sample of 445 post-primary schoolpupils in County Kilkenny, Ireland. Knowledge, attitudes and behaviour relevant to non-communicable diseases were examined using a questionnaire developed from a World Health Organization core questionnaire. The results showed that levels of adolescent alcohol and tobacco use were similar to those found in neighbouring European countries. Smoking and drinking increased during adolescence and were more prevalent in males. Physical activity decreased throughout adolescence and a high intake of "snack foods" was found. Health-related knowledge levels were high but were not related to behaviour.

Trends in mortality from cardiovascular diseases in Ireland.

Trends in mortality from cardiovascular diseases in Ireland from 1950 to 1986 are examined and compared with trends in some other developed countries. There was a decline in mortality from all causes which was greater in women than in men. Mortality from coronary heart disease (CHD) increased in males from 1953 to 1974 and has been declining slowly since. Mortality rates in men in Canada and the United States are now lower than those in Ireland and rates in Finnish men are approaching those in Ireland. CHD mortality in Irish males was sixth highest of developed countries in 1985. In women, CHD mortality declined in the 1950s, was stable in the 1960s and early 1970s and has been decreasing slowly since 1974. CHD mortality in Irish women was fifth highest of developed countries in 1985. Mortality rates from cerebrovascular disease have been falling in Irish women since 1958 and in men since 1969. The continuing high mortality from CHD in Ireland warrants the establishment of effective community prevention programmes.