P2X3 receptors participate in purinergic inhibition of gastrointestinal smooth muscle

The ATP analogue α,β-meATP is a potent relaxant of gastrointestinal smooth muscle, but its molecular target is uncertain inside the gut. α,β-meATP relaxed the carbachol-precontracted guinea-pig taenia coli in a concentration-dependent manner (EC50, 2.0 ± 0.1 μM). A luciferase-based assay confirmed that α,β-meATP solutions were minimally contaminated with ATP. α,β-meATP-evoked relaxations were inhibited by the competitive P2Y1 antagonist MRS2179 (pA2 = 5.36), but also by the competitive P2X3 antagonist, A-317491 (pA2 = 5.51). When MRS2179 and A-317491 were applied together, residual α,β-meATP responses converted from brief to prolonged relaxations. Sodium nitroprusside (a nitric oxide donor) also caused prolonged relaxations. Immunohistochemistry revealed that P2X3 receptors were present in myenteric ganglion cells and their varicose nerve terminals. The amplitude of α,β-meATP responses was not inhibited by TTX (NaV channel blocker) and ωCgTx (N-type CaV channel blocker). However, responses to α,β-meATP were inhibited by TEA (non-selective K+-channel blocker), indicating that relaxations involved opening K+-channels. The findings of this study are consistent with the conclusion that α,β-meATP stimulates Ca2+-permeable P2X3 receptors on varicose nerve terminals to release inhibitory nucleotides: 1) ATP and β-NAD release results in P2Y1-mediated brief relaxations; 2) another released transmitter (possibly NO) results in prolonged relaxations. Prejunctional P2X3 receptors represent a purinergic feed-forward mechanism to augment the action of inhibitory nerves on gut motility. This positive feed-forward mechanism may counter-balance the known negative feedback mechanism caused by adenosine and prejunctional A1 receptors on inhibitory motor nerves

Rehabilitation of the P2X5 receptor: a re-evaluation of structure and function

Of the extended family of ATP-gated P2X ion-channels, the P2X5 receptor has received comparatively little attention since first cloned over 25 years ago. Disinterest in studying this P2X subtype stems from two commonly held beliefs: (i) canonical human P2X5 is non-functional because the P2X5 subunit is truncated (hP2X5A, 422 aa) and missing the critical peptide sequence (22 aa) encoded by exon 10; (ii) rat and mouse P2X5 subunits are fully formed (455 aa) but the receptor is only weakly functional, and successive ATP responses rapidly run down in amplitude. However, newer studies have re-evaluated these notions. First, a low proportion (around 10%) of humans possess full-length P2X5 subunits (444 aa) and can form competent P2X5 receptors. Full-length P2X5 has been identified only in black Americans, but may occur in a wider population as more ethnicities are screened. Second, replacement of one of three amino acids in rat P2X5 subunits with corresponding residues in human P2X5 subunits (V67I, S191F, or F195H) significantly improves the responsiveness of rat P2X5 to ATP. Replaced residues exert an allosteric action on the left flipper, allowing the docking jaw for ATP to flex the lower body of the subunit and fully open the ion pore. This proposed action may drive the search for naturally occurring modulators which act allosterically on wildtype rat P2X5. This review collates the available information on the structure and function of human and rat P2X5 receptors, with the view to rehabilitating the reputation of these ATP-gated ion channels and stimulating future lines of research

Sensor node localisation using a stereo camera rig

In this paper, we use stereo vision processing techniques to detect and localise sensors used for monitoring simulated environmental events within an experimental sensor network testbed. Our sensor nodes communicate to the camera through patterns emitted by light emitting diodes (LEDs). Ultimately, we envisage the use of very low-cost, low-power, compact microcontroller-based sensing nodes that employ LED communication rather than power hungry RF to transmit data that is gathered via existing CCTV infrastructure. To facilitate our research, we have constructed a controlled environment where nodes and cameras can be deployed and potentially hazardous chemical or physical plumes can be introduced to simulate environmental pollution events in a controlled manner. In this paper we show how 3D spatial localisation of sensors becomes a straightforward task when a stereo camera rig is used rather than a more usual 2D CCTV camera

Maternal obesity has little effect on the immediate offspring but impacts on the next generation

Maternal obesity during pregnancy has been linked to an increased risk of obesity and cardiometabolic disease in the offspring, a phenomenon attributed to developmental programming. Programming effects may be transmissible across generations through both maternal and paternal inheritance, although the mechanisms remain unclear. Using a mouse model, we explored the effects of moderate maternal diet-induced obesity (DIO) on weight gain and glucose-insulin homeostasis in first-generation (F1) and second-generation offspring. DIO was associated with insulin resistance, hyperglycemia and dyslipidemia before pregnancy. Birth weight was reduced in female offspring of DIO mothers (by 6%, P = .039), and DIO offspring were heavier than controls at weaning (males by 47%, females by 27%), however there were no differences in glucose tolerance, plasma lipids, or hepatic gene expression at 6 months. Despite the relative lack of effects in the F1, we found clear fetal growth restriction and persistent metabolic changes in otherwise unmanipulated second-generation offspring with effects on birth weight, insulin levels, and hepatic gene expression that were transmitted through both maternal and paternal lines. This suggests that the consequences of the current dietary obesity epidemic may also have an impact on the descendants of obese individuals, even when the phenotype of the F1 appears largely unaffected

Evolution of an ancient protein function involved in organized multicellularity in animals.

To form and maintain organized tissues, multicellular organisms orient their mitotic spindles relative to neighboring cells. A molecular complex scaffolded by the GK protein-interaction domain (GKPID) mediates spindle orientation in diverse animal taxa by linking microtubule motor proteins to a marker protein on the cell cortex localized by external cues. Here we illuminate how this complex evolved and commandeered control of spindle orientation from a more ancient mechanism. The complex was assembled through a series of molecular exploitation events, one of which - the evolution of GKPID's capacity to bind the cortical marker protein - can be recapitulated by reintroducing a single historical substitution into the reconstructed ancestral GKPID. This change revealed and repurposed an ancient molecular surface that previously had a radically different function. We show how the physical simplicity of this binding interface enabled the evolution of a new protein function now essential to the biological complexity of many animals

A Cascade of Care of Patients with Hepatitis C Infection in a Rural State

Introduction: The substance misuse epidemic has fueled an increase in hepatitis C virus (HCV) infections. Despite the availability of sensitive screening and curative treatment, relatively few people are aware of their diagnosis and engaged in care. In this study, we aimed to identify local gaps in HCV care and inform strategies for improvement. Methods: In this retrospective study, we assessed adult patients seen at a tertiary care center from 2015 to 2019 and who were eligible for HCV screening based on recommendations from the Centers for Disease Control and Prevention. Inclusion criteria were birth from 1945 to 1965, long-term dialysis treatment, alanine aminotransferase greater than 35 U/L for 6 months or more, and/or a diagnosis of opioid use disorder (OUD), HIV/AIDS, or hepatitis B virus (HBV) infection. We summarized the HCV cascade of care with descriptive statistics and used logistic regression to identify factors associated with HCV screening. Results: We identified 4948 patients eligible for HCV screening, of whom 47% were female, 54% were male; 7% were Black, 83% were White, and 10% were Other/Unknown; and 87% were born between 1945 and 1965. Among the patients, 2791/4948 (56%) were screened and 124/2791 (4%) were identified to have chronic HCV infection, of whom 12/124 (10%) were linked to care, ever treated, and cured. Patients with HCV included 63/124 (51%) with OUD and 65/124 (52%) with HBV coinfection. All risk factors for HCV were independently associated with HCV screening, except OUD (aOR, 1.2; 95% CI, 0.9-1.6; P = .28). Discussion: We identified multiple gaps in the HCV cascade of care at our institution. Our findings, paired with data from the Veterans Health Administration and national research, indicate a need for more comprehensive strategies for HCV screening and intervention. Conclusions: Our findings will help to direct strategies for improving HCV detection and subsequent enrollment into care, particularly for patients with OUD

Is Your Neighborhood Designed to Support Physical Activity? A Brief Streetscape Audit Tool.

INTRODUCTION:Macro level built environment factors (eg, street connectivity, walkability) are correlated with physical activity. Less studied but more modifiable microscale elements of the environment (eg, crosswalks) may also affect physical activity, but short audit measures of microscale elements are needed to promote wider use. This study evaluated the relation of a 15-item neighborhood environment audit tool with a full version of the tool to assess neighborhood design on physical activity in 4 age groups. METHODS:From the 120-item Microscale Audit of Pedestrian Streetscapes (MAPS) measure of street design, sidewalks, and street crossings, we developed the 15-item version (MAPS-Mini) on the basis of associations with physical activity and attribute modifiability. As a sample of a likely walking route, MAPS-Mini was conducted on a 0.25-mile route from participant residences toward the nearest nonresidential destination for children (n = 758), adolescents (n = 897), younger adults (n = 1,655), and older adults (n = 367). Active transportation and leisure physical activity were measured with age-appropriate surveys, and accelerometers provided objective physical activity measures. Mixed-model regressions were conducted for each MAPS item and a total environment score, adjusted for demographics, participant clustering, and macrolevel walkability. RESULTS:Total scores of MAPS-Mini and the 120-item MAPS correlated at r = .85. Total microscale environment scores were significantly related to active transportation in all age groups. Items related to active transport in 3 age groups were presence of sidewalks, curb cuts, street lights, benches, and buffer between street and sidewalk. The total score was related to leisure physical activity and accelerometer measures only in children. CONCLUSION:The MAPS-Mini environment measure is short enough to be practical for use by community groups and planning agencies and is a valid substitute for the full version that is 8 times longer

Lattice effects observed in chaotic dynamics of experimental populations

Animals and many plants are counted in discrete units. The collection of possible values (state space) of population numbers is thus a nonnegative integer lattice. Despite this fact, many mathematical population models assume a continuum of system states. The complex dynamics, such as chaos, often displayed by such continuous-state models have stimulated much ecological research; yet discretestate models with bounded population size can display only cyclic behavior. Motivated by data from a population experiment, we compared the predictions of discrete-state and continuous-state population models. Neither the discrete- nor continuous-state models completely account for the data. Rather, the observed dynamics are explained by a stochastic blending of the chaotic dynamics predicted by the continuous-state model and the cyclic dynamics predicted by the discretestate models. We suggest that such lattice effects could be an important component of natural population fluctuations. The discovery that simple deterministic population models can display complex aperiodi

A Bioinformatics Method Identifies Prominent Off-targeted Transcripts in RNAi Screens

Because off-target effects hamper interpretation and validation of RNAi screens, we developed a bioinformatics method, Genome-wide Enrichment of Seed Sequence matches (GESS), to identify candidate off-targeted transcripts from direct analysis of primary screening data. GESS identified a prominent off-targeted transcript in several screens, including MAD2 in a screen for components of the spindle assembly checkpoint. We demonstrate how incorporation of the results of GESS analysis can enhance the validation rate in RNAi screens