Channel-mediated ATP release in the nervous system.

ATP is well established as a transmitter and modulator in the peripheral and central nervous system. While conventional exocytotic release of ATP at synapses occurs, this transmitter is unusual in also being released into the extracellular space via large-pored plasma membrane channels. This review considers the channels that are known to be permeable to ATP and some of the functions of channel-mediated ATP release. While the possibility of ATP release via channels mediating volume transmission has been known for some time, localised ATP release via channels at specialised synapses made by taste cells to the afferent nerve has recently been documented in taste buds. This raises the prospect that "channel synapses" may occur in other contexts. However, volume transmission and channel synapses are not necessarily mutually exclusive. We suggest that certain glial cells in the brain stem and hypothalamus, which possess long processes and are known to release ATP, may be candidates for both modes of ATP release -channel-mediated volume transmission in the region of their somata and more localised transmission possibly via either conventional or channel synapses from their processes at distal targets. Finally, we consider the different characteristics of vesicular and channel synapses and suggest that channel synapses may be advantageous in requiring less energy than their conventional vesicular counterparts. This article is part of the Special Issue on "Purinergic Signaling: 50 years".

X-linked Charcot Marie Tooth mutations alter CO2 sensitivity of connexin32 hemichannels.

Connexin32 (Cx32) is expressed in myelinating Schwann cells. It forms both reflexive gap junctions, to facilitate transfer of molecules from the outer to the inner myelin layers and hemichannels at the paranode to permit action potential-evoked release of ATP into the extracellular space. Loss of function mutations in Cx32 cause X-linked Charcot Marie Tooth disease (CMTX), a slowly developing peripheral neuropathy. The mechanistic links between Cx32 mutations and CMTX are not well understood. As Cx32 hemichannels can be opened by increases in PCO2, we have examined whether CMTX mutations alter this CO2 sensitivity. By using Ca2+ imaging, dye loading and genetically encoded ATP sensors to measure ATP release, we have found 5 CMTX mutations that abolish the CO2 sensitivity of Cx32 hemichannels (A88D, 111-116 Del, C179Y, E102G, V139M). Others cause a partial loss (L56F, R220Stop, and R15W). Some CMTX mutations have no apparent effect on CO2 sensitivity (R15Q, L9F, G12S, V13L, V84I, W133R). The mutation R15W alters multiple additional aspects of hemichannel function including Ca2+ and ATP permeability. The mutations that abolish CO2 sensitivity are transdominant and abolish CO2 sensitivity of co-expressed Cx32WT. We have shown that Schwannoma RT4 D6P2T cells can release ATP in response to elevated PCO2 via the opening of Cx32. This is consistent with the hypothesis that the CO2 sensitivity of Cx32 may be important for maintenance of healthy myelin. Our data, showing a transdominant effect of certain CMTX mutations on CO2 sensitivity, may need to be taken into account in any future gene therapies for this condition.

Belowground mechanism reveals climate change impacts on invasive clonal plant establishment.

Climate change and disturbance can alter invasion success of clonal plants by differentially affecting the clonal traits influencing their establishment as young plants. Clonal traits related to the vegetative reproduction of native Pascopyrum smithii and non-native Bromus inermis grass seedlings were evaluated under altered precipitation frequencies and a single grazing event. Pascopyrum smithii maintained similar vegetative reproduction under three simulated precipitation frequencies whereas B. inermis vegetative reproduction declined as precipitation became more intermittent. Vegetative reproduction of the non-native B. inermis was greater than the native P. smithii under all simulated precipitation frequencies except the most intermittent scenario. A single grazing event did not affect either species' response to intra-annual precipitation variability but did slightly reduce their clonal growth and increase their bud dormancy. In young plants, clonal traits of the invasive grass favored its superior expansion and population growth compared to the native grass except under the most severe climate change scenario. Grassland restoration using native P. smithii seeds would be successful in most years due to its resilient clonal growth in a changing climate. Clonal infrastructure development in young plants is critical to clonal plant establishment and persistence in a changing climate and under disturbed conditions.

Chlamydomonas ARMC2/PF27 is an obligate cargo adapter for intraflagellar transport of radial spokes.

Intraflagellar transport (IFT) carries proteins into flagella but how IFT trains interact with the large number of diverse proteins required to assemble flagella remains largely unknown. Here, we show that IFT of radial spokes in Chlamydomonas requires ARMC2/PF27, a conserved armadillo repeat protein associated with male infertility and reduced lung function. Chlamydomonas ARMC2 was highly enriched in growing flagella and tagged ARMC2 and the spoke protein RSP3 co-migrated on anterograde trains. In contrast, a cargo and an adapter of inner and outer dynein arms moved independently of ARMC2, indicating that unrelated cargoes distribute stochastically onto the IFT trains. After concomitant unloading at the flagellar tip, RSP3 attached to the axoneme whereas ARMC2 diffused back to the cell body. In armc2/pf27 mutants, IFT of radial spokes was abolished and the presence of radial spokes was limited to the proximal region of flagella. We conclude that ARMC2 is a cargo adapter required for IFT of radial spokes to ensure their assembly along flagella. ARMC2 belongs to a growing class of cargo-specific adapters that enable flagellar transport of preassembled axonemal substructures by IFT.

Field Experiments and Meta-analysis Reveal Wetland Vegetation as a Crucial Element in the Coastal Protection Paradigm.

Increasing rates of sea-level rise and wave action threaten coastal populations. Defense of shorelines by protection and restoration of wetlands has been invoked as a win-win strategy for humans and nature, yet evidence from field experiments supporting the wetland protection function is uncommon, as is the understanding of its context dependency. Here we provide evidence from field manipulations showing that the loss of wetland vegetation, regardless of disturbance size, increases the rate of erosion on wave-stressed shorelines. Vegetation removal (simulated disturbance) along the edge of salt marshes reveals that loss of wetland plants elevates the rate of lateral erosion and that extensive root systems, rather than aboveground biomass, are primarily responsible for protection against edge erosion in marshes. Meta-analysis further shows that disturbances that generate plant die-off on salt marsh edges generally hasten edge erosion in coastal marshes and that the erosion protection function of wetlands relates more to lateral than vertical edge-erosional processes and is positively correlated with the amount of belowground plant biomass lost. Collectively, our findings substantiate a coastal protection paradigm that incorporates preservation of shoreline vegetation, illuminate key context dependencies in this theory, and highlight local disturbances (e.g., oil spills) that kill wetland plants as agents that can accelerate coastal erosion.

Secondary invasion re-redefined: The distinction between invader-facilitated and invader-contingent invasions as subclasses of secondary invasion.

Linked Article: https://doi.org/10.1002/ece3.3964.

Relative Influence of Trans-Pacific and Regional Atmospheric Transport of PAHs in the Pacific Northwest, U.S.

The relative influences of trans-Pacific and regional atmospheric transport on measured concentrations of polycyclic aromatic hydrocarbons (PAHs), PAH derivatives (nitro- (NPAH) and oxy-(OPAH)), organic carbon (OC), and particulate matter (PM) less than 2.5 µm in diameter (PM2.5) were investigated in the Pacific Northwest, U.S. in 2010-2011. Ambient high volume PM2.5 air samples were collected at two sites in the Pacific Northwest: (1.) Mount Bachelor Observatory (MBO) in the Oregon Cascade Range (2763 m above sea level (asl)) and 2.) Confederated Tribes of the Umatilla Indian Reservation (CTUIR) in the Columbia River Gorge (CRG) (954 m asl). At MBO, the 1,8-dinitropyrene concentration was significantly positively correlated with the time a sampled air mass spent over Asia, suggesting that this NPAH may be a good marker for trans-Pacific atmospheric transport. At CTUIR, NOx, CO2, and SO2 emissions from a 585 MW coal fired power plant, in Boardman OR, were found to be significantly positively correlated with PAH, OPAH, NPAH, OC, and PM2.5 concentrations. By comparing the Boardman Plant operational time frames when the plant was operating to when it was shut down, the plant was found to contribute a large percentage of the measured PAH (67%), NPAH (91%), OPAH (54%), PM2.5 (39%), and OC (38%) concentrations at CTUIR and the CRG prior to Spring 2011 and likely masked trans-Pacific atmospheric transport events to the CRG. Upgrades installed to the Boardman Plant in the spring of 2011 dramatically reduced the plant's contribution to PAH and OPAH concentrations (by ∼72% and ∼40%, respectively) at CTUIR and the CRG, but not NPAH, PM2.5 or OC concentrations.

Clinical impression and ascites appearance do not rule out bacterial peritonitis.

BACKGROUND: Previous research has demonstrated that physician clinical suspicion, determined without assessing fluid appearance, is not adequate to rule out spontaneous bacterial peritonitis (SBP) without fluid testing. STUDY OBJECTIVE: To determine the sensitivity of physician clinical suspicion, including a bedside assessment of fluid appearance, in the detection of SBP in Emergency Department (ED) patients undergoing paracentesis. METHODS: We conducted a prospective, observational study of ED patients with ascites undergoing paracentesis at three academic facilities. The enrolling physician recorded the clinical suspicion of SBP ("none," "low," "moderate," or "high"), and ascites appearance ("clear," "hazy," "cloudy," or "bloody"). SBP was defined as an absolute neutrophil count ≥ 250 cells/mm(3), or culture pathogen growth. We defined "clear" ascites fluid as negative for SBP, and "hazy," "cloudy," or "bloody" as positive. A physician clinical suspicion of "none" or "low" was considered negative for SBP, and an assessment of "moderate" or "high" was considered positive. The primary outcome measure was sensitivity of physician clinical impression and ascites appearance for SBP. RESULTS: There were 348 cases enrolled, with SBP diagnosed in 43 (12%). Physician clinical suspicion had a sensitivity of 42% (95% confidence interval [CI] 29-55%) for the detection of SBP. Fluid appearance had a sensitivity of 72% (95% CI 58-83%). CONCLUSION: Physician clinical impression, which included an assessment of fluid appearance, had poor sensitivity for the detection of SBP and cannot be used to exclude the diagnosis. Routine laboratory fluid analysis is indicated after ED paracentesis, even in patients considered to have a low degree of suspicion for SBP.

E-portfolio competency metadata: pilot study for a call to action.

The six competency domains required by the Accreditation Council for Graduate Medical Education (ACGME) have led to a proliferation of measurement tools, assessment methods, and all forms of data from paper to electronic. The need exists to develop a standardized electronic (e)-portfolio to provide the aggregate data to improve education and patient care. This process requires a sound methodology using XML metadata to allow portability of e-portfolio data. We surveyed publicly available metadata and developed an e-portfolio system for the Henry Ford Hospital General Surgery Residency Program. Based on our implementation of e-portfolios for 70 physicians, we call upon the ACGME, the Residency Review Committees, and the American Board of Medical Specialties to establish a method to formalize and develop a standard for residency competency metadata. Using an approach similar to that of our study can streamline data and lead to improved medical education and ultimately better patient care.

Top predators suppress rather than facilitate plants in a trait-mediated tri-trophic cascade.

Classical ecological theory states that in tri-trophic systems, predators indirectly facilitate plants by reducing herbivore densities through consumption, while more recent work has revealed that predators can generate the same positive effect on plants non-consumptively by inducing changes in herbivore traits (e.g. feeding rates). Based on observations in US salt marshes dominated by vast monocultures of cordgrass, we hypothesized that sit-and-wait substrate-dwelling predators (crabs) could actually strengthen per capita impacts of potent grazers (snails), by non-consumptively inducing a vertical habitat shift of snails to their predation refuge high on canopy leaves that are vulnerable to grazing. A two-month field experiment supported this hypothesis, revealing that predators non-consumptively increased the mean climbing height of snails on grasses, increased grazing damage per leaf and ultimately suppressed cordgrass biomass, relative to controls. While seemingly counterintuitive, our results can be explained by (i) the vulnerability of refuge resources to grazing, and (ii) universal traits that drove the vertical habitat shift--i.e. relative habitat domains of predator and prey, and the hunting mode of the top predator. These results underline the fact that not only should we continue to incorporate non-consumptive effects into our understanding of top-down predator impacts, but we should do so in a spatially explicit manner.

An unusual cause of pelvic pain and fever: periurethral abscess from an infected urethral diverticulum.

BACKGROUND: The evaluation of patients with pelvic pain is a common task for emergency physicians. Accurate diagnosis of the underlying cause of pelvic pain in women is often difficult given the diversity of pathology that can generate pelvic pain. OBJECTIVE: To report a rare but clinically significant cause of acute pelvic pain in women. CASE REPORT: We describe a rare case of a periurethral abscess secondary to an infected urethral diverticulum in a 41-year-old woman. She presented to our Emergency Department with pelvic pain, dysuria, and fever of 4 days duration. CONCLUSION: Physical examination, imaging studies, and surgery aided in confirming the diagnosis of a periurethral abscess.

Physician clinical impression does not rule out spontaneous bacterial peritonitis in patients undergoing emergency department paracentesis.

STUDY OBJECTIVE: We determine whether clinical characteristics and physician assessment are useful in the exclusion of spontaneous bacterial peritonitis in emergency department (ED) patients with ascites requiring paracentesis. METHODS: We conducted a prospective, observational study of ED patients with ascites undergoing paracentesis. Predefined clinical characteristics including historical features and ED vital signs were recorded. Each patient was assessed by 2 separate, blinded physicians for severity of abdominal tenderness and overall clinical suspicion for spontaneous bacterial peritonitis. The primary outcome measures were sensitivity, specificity, and likelihood ratios (LR) of the individual clinical characteristics and the physician assessments. Spontaneous bacterial peritonitis was defined by absolute neutrophil count greater than 250 cells/mm(3) or positive fluid culture result. RESULTS: There were 285 separate physician assessments in 144 patients enrolled with complete data. Spontaneous bacterial peritonitis was diagnosed in 17 (11.8%) patients. Physician clinical impression had a sensitivity of 76% (95% confidence interval [CI] 62% to 91%) and specificity of 34% (95% CI 28% to 40%) for the detection of spontaneous bacterial peritonitis. The lowest negative LR was associated with the presence of any abdominal pain or tenderness (negative LR=0.4); however, the presence of pain/tenderness was also observed in 85% of patients without spontaneous bacterial peritonitis. Six patients (4.2%) with spontaneous bacterial peritonitis had at least 1 physician assessment of little to no risk for spontaneous bacterial peritonitis, and 3 of the 6 subsequently died during their hospitalization. CONCLUSION: Clinical characteristics and physician assessment were insufficient in the diagnosis or exclusion of spontaneous bacterial peritonitis in the ED patient undergoing diagnostic or therapeutic paracentesis. This finding supports routine laboratory fluid analysis after ED paracentesis.

An unusual complication of epididymo-orchitis: scrotal pyocele extending into the inguinal canal mimicking a strangulated inguinal hernia.

We report a case of a paraplegic patient with an indwelling Foley catheter and epididymo-orchitis leading to a scrotal pyocele. This case is unique in that the patient's physical examination revealed both inguinal and scrotal abnormalities that mimicked a strangulated inguinal hernia. Imaging studies and surgical exploration helped elucidate the exact diagnosis. Although rare, this entity requires emergent surgical consultation. We will also discuss the etiology, pathophysiology, and treatment of scrotal abscesses.