Mechanistic basis of the dynamic response of TWIK1 ionic selectivity to pH.

Highly selective for K+ at neutral pH, the TWIK1 channel becomes permeable to Na+ upon acidification. Using molecular dynamics simulations, we identify a network of residues involved in this unique property. Between the open and closed states previously observed by electron microscopy, molecular dynamics simulations show that the channel undergoes conformational changes between pH 7.5-6 involving residues His122, Glu235, Lys246 and Phe109. A complex network of interactions surrounding the selectivity filter at high pH transforms into a simple set of stronger interactions at low pH. In particular, His122 protonated by acidification moves away from Lys246 and engages in a salt bridge with Glu235. In addition, stacking interactions between Phe109 and His122, which stabilize the selectivity filter in its K+-selective state at high pH, disappear upon acidification. This leads to dissociation of the Phe109 aromatic side chain from this network, resulting in the Na+-permeable conformation of the channel.

Impact of deep learning image reconstruction on volumetric accuracy and image quality of pulmonary nodules with different morphologies in low-dose CT.

BACKGROUND: This study systematically compares the impact of innovative deep learning image reconstruction (DLIR, TrueFidelity) to conventionally used iterative reconstruction (IR) on nodule volumetry and subjective image quality (IQ) at highly reduced radiation doses. This is essential in the context of low-dose CT lung cancer screening where accurate volumetry and characterization of pulmonary nodules in repeated CT scanning are indispensable. MATERIALS AND METHODS: A standardized CT dataset was established using an anthropomorphic chest phantom (Lungman, Kyoto Kaguku Inc., Kyoto, Japan) containing a set of 3D-printed lung nodules including six diameters (4 to 9 mm) and three morphology classes (lobular, spiculated, smooth), with an established ground truth. Images were acquired at varying radiation doses (6.04, 3.03, 1.54, 0.77, 0.41 and 0.20 mGy) and reconstructed with combinations of reconstruction kernels (soft and hard kernel) and reconstruction algorithms (ASIR-V and DLIR at low, medium and high strength). Semi-automatic volumetry measurements and subjective image quality scores recorded by five radiologists were analyzed with multiple linear regression and mixed-effect ordinal logistic regression models. RESULTS: Volumetric errors of nodules imaged with DLIR are up to 50% lower compared to ASIR-V, especially at radiation doses below 1 mGy and when reconstructed with a hard kernel. Also, across all nodule diameters and morphologies, volumetric errors are commonly lower with DLIR. Furthermore, DLIR renders higher subjective IQ, especially at the sub-mGy doses. Radiologists were up to nine times more likely to score the highest IQ-score to these images compared to those reconstructed with ASIR-V. Lung nodules with irregular margins and small diameters also had an increased likelihood (up to five times more likely) to be ascribed the best IQ scores when reconstructed with DLIR. CONCLUSION: We observed that DLIR performs as good as or even outperforms conventionally used reconstruction algorithms in terms of volumetric accuracy and subjective IQ of nodules in an anthropomorphic chest phantom. As such, DLIR potentially allows to lower the radiation dose to participants of lung cancer screening without compromising accurate measurement and characterization of lung nodules.

Alkaline-sensitive two-pore domain potassium channels form functional heteromers in pancreatic ß-cells.

Two-pore domain K+ channels (K2P channels), active as dimers, produce inhibitory currents regulated by a variety of stimuli. Among them, TWIK1-related alkalinization-activated K+ channel 1 (TALK1), TWIK1-related alkalinization-activated K+ channel 2 (TALK2), and TWIK1-related acid-sensitive K+ channel 2 (TASK2) form a subfamily of structurally related K2P channels stimulated by extracellular alkalosis. The human genes encoding these proteins are clustered at chromosomal region 6p21 and coexpressed in multiple tissues, including the pancreas. The question whether these channels form functional heteromers remained open. By analyzing single-cell transcriptomic data, we show that these channels are coexpressed in insulin-secreting pancreatic ß-cells. Using in situ proximity ligation assay and electrophysiology, we show that they form functional heterodimers both upon heterologous expression and under native conditions in human pancreatic ß-cells. We demonstrate that heteromerization of TALK2 with TALK1 or with TASK2 endows TALK2 with sensitivity to extracellular alkalosis in the physiological range. We further show that the association of TASK2 with TALK1 and TALK2 increases their unitary conductance. These results provide a new example of heteromerization in the K2P channel family expanding the range of the potential physiological and pathophysiological roles of TALK1/TALK2/TASK2 channels, not only in insulin-secreting cells but also in the many other tissues in which they are coexpressed.

Corrigendum: Convergence of Multiple Stimuli to a Single Gate in TREK1 and TRAAK Potassium Channels.

[This corrects the article DOI: 10.3389/fphar.2021.755826.].

Piezo1 and Piezo2 foster mechanical gating of K2P channels.

Mechanoelectrical transduction is mediated by the opening of different types of force-sensitive ion channels, including Piezo1/2 and the TREK/TRAAK K2P channels. Piezo1 curves the membrane locally into an inverted dome that reversibly flattens in response to force application. Moreover, Piezo1 forms numerous preferential interactions with various membrane lipids, including cholesterol. Whether this structural architecture influences the functionality of neighboring membrane proteins is unknown. Here, we show that Piezo1/2 increase TREK/TRAAK current amplitude, slow down activation/deactivation, and remove inactivation upon mechanical stimulation. These findings are consistent with a mechanism whereby Piezo1/2 cause a local depletion of membrane cholesterol associated with a prestress of TREK/TRAAK channels. This regulation occurs in mouse fibroblasts between endogenous Piezo1 and TREK-1/2, both channel types acting in concert to delay wound healing. In conclusion, we demonstrate a community effect between different structural and functional classes of mechanosensitive ion channels.

Applying a One Health Approach in Global Health and Medicine: Enhancing Involvement of Medical Schools and Global Health Centers.

Background: Multidisciplinary and multisectoral approaches such as One Health and related concepts (e.g., Planetary Health, EcoHealth) offer opportunities for synergistic expertise to address complex health threats. The connections between humans, animals, and the environment necessitate collaboration among sectors to comprehensively understand and reduce risks and consequences on health and wellbeing. One Health approaches are increasingly emphasized for national and international plans and strategies related to zoonotic diseases, food safety, antimicrobial resistance, and climate change, but to date, the possible applications in clinical practice and benefits impacting human health are largely missing. Methods: In 2018 the "Application of the One Health Approach to Global Health Centers" conference held at the Albert Einstein College of Medicine convened experts involved in One Health policy and practice. The conference examined issues relevant to One Health approaches, sharing examples of challenges and successes to guide application to medical school curricula and clinical practice for human health. This paper presents a synthesis of conference proceedings, framed around objectives identified from presentations and audience feedback. Findings and Recommendations: The following objectives provide opportunities for One Health involvement and benefits for medical schools and global health centers by: 1) Improving One Health resource sharing in global health and medical education; 2) Creating pathways for information flow in clinical medicine and global health practice; 3) Developing innovative partnerships for improved health sector outcomes; and 4) Informing and empowering health through public outreach. These objectives can leverage existing resources to deliver value to additional settings and stakeholders through resource efficiency, more holistic and effective service delivery, and greater ability to manage determinants of poor health status. We encourage medical and global health educators, practitioners, and students to explore entry points where One Health can add value to their work from local to global scale.

PULMONARY NODULE DETECTION IN CHEST CT USING A DEEP LEARNING-BASED RECONSTRUCTION ALGORITHM.

This study's aim was to assess whether deep learning image reconstruction (DLIR) techniques are non-inferior to ASIR-V for the clinical task of pulmonary nodule detection in chest computed tomography. Up to 6 (range 3-6, mean 4.2) artificial lung nodules (diameter: 3, 5, 8 mm; density: -800, -630, +100 HU) were inserted at different locations in the Kyoto Kagaku Lungman phantom. In total, 16 configurations (10 abnormal, 6 normal) were scanned at 7.6, 3, 1.6 and 0.38 mGy CTDIvol (respectively 0, 60, 80 and 95% dose reduction). Images were reconstructed using 50% ASIR-V and a deep learning-based algorithm with low (DL-L), medium (DL-M) and high (DL-H) strength. Four chest radiologists evaluated 256 series by locating and scoring nodules on a five-point scale. No statistically significant difference was found among the reconstruction algorithms (p = 0.987, average across readers AUC: 0.555, 0.561, 0.557, 0.558 for ASIR-V, DL-L, DL-M, DL-H).

Physiological roles of heteromerization: focus on the two-pore domain potassium channels.

Potassium channels form the largest family of ion channels with more than 80 members involved in cell excitability and signalling. Most of them exist as homomeric channels, whereas specific conditions are required to obtain heteromeric channels. It is well established that heteromerization of voltage-gated and inward rectifier potassium channels affects their function, increasing the diversity of the native potassium currents. For potassium channels with two pore domains (K2P ), homomerization has long been considered the rule, their polymodal regulation by a wide diversity of physical and chemical stimuli being responsible for the adaptation of the leak potassium currents to cellular needs. This view has recently evolved with the accumulation of evidence of heteromerization between different K2P subunits. Several functional intragroup and intergroup heteromers have recently been identified, which contribute to the functional heterogeneity of this family. K2P heteromerization is involved in the modulation of channel expression and trafficking, promoting functional and signalling diversity. As illustrated in the Abstract Figure, heteromerization of TREK1 and TRAAK provides the cell with more possibilities of regulation. It is becoming increasingly evident that K2P heteromers contribute to important physiological functions including neuronal and cardiac excitability. Since heteromerization also affects the pharmacology of K2P channels, this understanding helps to establish K2P heteromers as new therapeutic targets for physiopathological conditions.

Distinct classes of potassium channels fused to GPCRs as electrical signaling biosensors.

Ligand-gated ion channels (LGICs) are natural biosensors generating electrical signals in response to the binding of specific ligands. Creating de novo LGICs for biosensing applications is technically challenging. We have previously designed modified LGICs by linking G protein-coupled receptors (GPCRs) to the Kir6.2 channel. In this article, we extrapolate these design concepts to other channels with different structures and oligomeric states, namely a tetrameric viral Kcv channel and the dimeric mouse TREK-1 channel. After precise engineering of the linker regions, the two ion channels were successfully regulated by a GPCR fused to their N-terminal domain. Two-electrode voltage-clamp recordings showed that Kcv and mTREK-1 fusions were inhibited and activated by GPCR agonists, respectively, and antagonists abolished both effects. Thus, dissimilar ion channels can be allosterically regulated through their N-terminal domains, suggesting that this is a generalizable approach for ion channel engineering.

Acromioplasty during repair of rotator cuff tears removes only half of the impinging acromial bone.

BACKGROUND: To date, there is no consensus on when and how to perform acromioplasty during rotator cuff repair (RCR). We aimed to determine the volume of impinging bone removed during acromioplasty and whether it influences postoperative range of motion (ROM) and clinical scores after RCR. METHODS: Preoperative and postoperative computed tomography scans of 57 shoulders that underwent RCR were used to reconstruct scapula models to simulate volumes of impinging acromial bone preoperatively and then compare them to the volumes of bone resected postoperatively to calculate the proportions of desired (ideal) vs. unnecessary (excess) resections. All patients were evaluated preoperatively and at 6 months to assess ROM and functional scores. RESULTS: The volume of impinging bone identified was 3.5 ± 2.3 cm3, of which 1.6 ± 1.2 cm3 (50% ± 27%) was removed during acromioplasty. The volume of impinging bone identified was not correlated with preoperative critical shoulder angle (r = 0.025, P = .853), nor with glenoid inclination (r = -0.024, P = .857). The volume of bone removed was 3.7 ± 2.2 cm3, of which 2.1 ± 1.6 cm3 (53% ± 24%) were unnecessary resections. Multivariable analyses revealed that more extensive removal of impinging bone significantly improved internal rotation with the arm at 90° of abduction (beta, 27.5, P = .048) but did not affect other shoulder movements or clinical scores. CONCLUSIONS: Acromioplasty removed only 50% of the estimated volume of impinging acromial bone. More extensive removal of impinging bone significantly improved internal rotation with the arm at 90° of abduction.

Guided versus freehand acromioplasty during rotator cuff repair. A randomized prospective study.

INTRODUCTION: There is no consensus on how to perform acromioplasty, particularly regarding the level and extent of bone resection, which depend on scapular and humeral morphologies. HYPOTHESIS: We aimed to determine whether computer-assisted acromioplasty planning helps surgeons remove impinging bone, reduce unnecessary resections, and improve short-term outcomes of rotator cuff tears (RCR). PATIENTS AND METHODS: We randomized 64 patients undergoing RCR of full-thickness supraspinatus tears into two groups: 'guided acromioplasty' (GA) and 'freehand acromioplasty' (FA). The pre- and post-operative scapula models were reconstructed using computed-tomography scans to quantify impinging bone removal, unnecessary bone resections, and identify zones of acromial bone removal. All patients were evaluated preoperatively and at 6 months to assess their range of motion (ROM), functional scores and tendon integrity using ultrasound. RESULTS: The two groups did not differ in demographics, clinical or morphologic characteristics. Compared to FA, GA tended to lower impinging bone removal (55±26% vs. 43±27%, p=0.087) and to increase unnecessary resection of the total bone removed (49±22% vs. 57±27%, p=0.248). GA resulted in significant anterior under-resection, while FA resulted in significant medial over-resection. Clinical outcomes and ROM improved significantly for all patients, except for internal rotation in the GA group. There were no other significant differences between the two groups, neither in terms of post-operative scores nor in terms of clinical net improvements, nor tendon repair integrity. CONCLUSIONS: This computer-assisted planning for acromioplasty during RCR proved no benefits in terms of bone removal, tendon healing, or clinical outcomes. Nonetheless such planning tools could help less experienced surgeons improve the efficacy of acromioplasty. LEVEL OF PROOF: I, Randomized controlled trial (Therapeutic study).

Selectivity filter instability dominates the low intrinsic activity of the TWIK-1 K2P K+ channel.

Two-pore domain K+ (K2P) channels have many important physiological functions. However, the functional properties of the TWIK-1 (K2P1.1/KCNK1) K2P channel remain poorly characterized because heterologous expression of this ion channel yields only very low levels of functional activity. Several underlying reasons have been proposed, including TWIK-1 retention in intracellular organelles, inhibition by posttranslational sumoylation, a hydrophobic barrier within the pore, and a low open probability of the selectivity filter (SF) gate. By evaluating these potential mechanisms, we found that the latter dominates the low intrinsic functional activity of TWIK-1. Investigating this further, we observed that the low activity of the SF gate appears to arise from the inefficiency of K+ in stabilizing an active (i.e. conductive) SF conformation. In contrast, other permeant ion species, such as Rb+, NH4+, and Cs+, strongly promoted a pH-dependent activated conformation. Furthermore, many K2P channels are activated by membrane depolarization via an SF-mediated gating mechanism, but we found here that only very strong nonphysiological depolarization produces voltage-dependent activation of heterologously expressed TWIK-1. Remarkably, we also observed that TWIK-1 Rb+ currents are potently inhibited by intracellular K+ (IC50 = 2.8 mm). We conclude that TWIK-1 displays unique SF gating properties among the family of K2P channels. In particular, the apparent instability of the conductive conformation of the TWIK-1 SF in the presence of K+ appears to dominate the low levels of intrinsic functional activity observed when the channel is expressed at the cell surface.

Electron attachment to hexafluoropropylene oxide (HFPO).

We probe the electron attachment in hexafluoropropylene oxide (HFPO), C3F6O, a gas widely used in plasma technologies. We determine the absolute electron attachment cross section using two completely different experimental approaches: (i) a crossed-beam experiment at single collision conditions (local pressures of 5 × 10-4 mbar) and (ii) a pulsed Townsend experiment at pressures of 20-100 mbar. In the latter method, the cross sections are unfolded from the electron attachment rate coefficients. The cross sections derived independently by the two methods are in very good agreement. We additionally discuss the dissociative electron attachment fragmentation patterns and their role in the radical production in industrial HFPO plasmas.

Institutionalizing One Health: From Assessment to Action.

A One Health approach is critical to strengthening health security at country, regional, and global levels. However, operationally its uptake remains limited. Recent momentum in assessing capacity to effectively prevent, detect, and respond to disease threats has resulted in identification of gaps that require dedicated action. This article highlights relevant tools, standards, and guidance to assist countries and institutions in meeting the collective vision articulated at the 2018 Prince Mahidol Award Conference on "Making the World Safe from the Threats of Emerging Infectious Diseases." Taking stock of assessment findings, resources, priorities, and implementation initiatives across human and animal health, environment and disaster risk reduction sectors can help expand participation in global health security, target risk drivers, and form synergies for collective action and shared gains for both emerging and endemic disease challenges. In addition to health security gains, a multisectoral, One Health approach can drive benefits for wider health sector and global development goals.

Antagonistic Effect of a Cytoplasmic Domain on the Basal Activity of Polymodal Potassium Channels.

TREK/TRAAK channels are polymodal K+ channels that convert very diverse stimuli, including bioactive lipids, mechanical stretch and temperature, into electrical signals. The nature of the structural changes that regulate their activity remains an open question. Here, we show that a cytoplasmic domain (the proximal C-ter domain, pCt) exerts antagonistic effects in TREK1 and TRAAK. In basal conditions, pCt favors activity in TREK1 whereas it impairs TRAAK activity. Using the conformation-dependent binding of fluoxetine, we show that TREK1 and TRAAK conformations at rest are different, and under the influence of pCt. Finally, we show that depleting PIP2 in live cells has a more pronounced inhibitory effect on TREK1 than on TRAAK. This differential regulation of TREK1 and TRAAK is related to a previously unrecognized PIP2-binding site (R329, R330, and R331) present within TREK1 pCt, but not in TRAAK pCt. Collectively, these new data point out pCt as a major regulatory domain of these channels and suggest that the binding of PIP2 to the pCt of TREK1 results in the stabilization of the conductive conformation in basal conditions.

Detailed precision and accuracy analysis of swarm parameters from a pulsed Townsend experiment.

A newly built pulsed Townsend experimental setup which allows one to measure both electron and ion currents is presented. The principle of pulsed Townsend measurements itself is well established to obtain swarm parameters such as the effective ionization rate coefficient, the density-reduced mobility, and the density-normalized longitudinal diffusion coefficient. The main novelty of the present contribution is a detailed and comprehensive analysis of the entire measurement and evaluation chain with respect to accuracy, precision, and reproducibility. The influence of the input parameters (gap distance, applied voltage, measured pressure, and temperature) is analyzed in detail. An overall accuracy of ±0.5% in the density reduced electric field (E/N) is achieved, which is close to the theoretically possible limit using the chosen components. The precision of the experimental results is higher than the accuracy. Through an extensive measurement campaign, the repeatability of our measurements proved to be high and similar to the precision. The reproducibility of results at identical (E/N) is similar to the precision for different distances but decreases for varying pressures. For benchmark purposes, measurements for Ar, CO2, and N2 are presented and compared with our previous experimental setup, simulations, and other experimental references.

[Prevalence, Risk Factors and Diagnostics of Hearing Impairment in Preterm Infants]. / Prävalenz, Risikofaktoren und Diagnostik von Hörstörungen bei Frühgeborenen.

Introduction: The preterm birth is clearly associated with increased risk of developing congenital hearing impairment. Therefore, special attention must be paid to the postnatal control of auditory function in all preterm infants. The present work investigates if the latest scientific findings regarding prevalence, clinical diagnostics, therapy and risk factors of hearing impairment in premature infants are regularly implemented in daily practice. Methods: At the department of phoniatrics and pediatric audiology of the University Hospital of Magdeburg, the treatment data of 126 preterm children born between 2006 and 2011 were evaluated retrospectively. The additional analysis of all records available at the screening center (n=67 640) covering this period enables drawing conclusions on the total number and prevalence of hearing impairment in preterm infants in Saxony-Anhalt. Results: Almost all premature babies, like mature newborns, underwent postnatal hearing screening of both ears. The data analysis shows that the practical implementation often does not comply with the guideline of the G-BA (Gemeinsamer Bundesausschuss) in all details. For example, the recommended screening method for preterm infants (AABR) or the screening and treatment timing are not always applied in accordance with the guidelines of the G-BA. Discussion: Assessment of the practical implementation of universal newborn hearing screening was planned at the time of the introduction of the hearing screening program by the G-BA. As a part of this investigation, the practical care of vulnerable groups such as preterm infants must be given special attention. Based on the collected data, the diagnostics and therapy should be unified. Regardless of the maternity clinic where the infants were born, there should be the same opportunity for early diagnosis and thus for prognostically better treatment of congenital hearing impairment. Rapid postnatal fitting with hearing aid can stimulate the maturation of the central auditory system and potentially help to avoid problems of hearing and speech development.

Development of the First Two-Pore Domain Potassium Channel TWIK-Related K+ Channel 1-Selective Agonist Possessing in Vivo Antinociceptive Activity.

The TWIK-related K+ channel, TREK-1, has recently emerged as an attractive therapeutic target for the development of a novel class of analgesic drugs, suggesting that activation of TREK-1 could result in pain inhibition. Here, we report the synthesis of a series of substituted acrylic acids (1-54) based on our previous work with caffeate esters. The analogues were evaluated for their ability to modulate TREK-1 channel by electrophysiology and for their in vivo antinociceptive activity (acetic acid-induced writhing and hot plate assays), leading to the identification of a series of novel molecules able to activate TREK-1 and displaying potent antinociceptive activity in vivo. Furyl analogue 36 is the most promising of the series.

Substance exposure and helmet use in all-terrain vehicle accidents: Nine years of experience at a level 1 trauma center.

INTRODUCTION: The surging popularity of all-terrain vehicles (ATV) in the United States has caused an "epidemic of injuries and mortality." The U.S. Consumer Product Safety Commission reported 99,600 injuries and 426 fatalities from ATV accidents in 2013. The aim of this study was to examine the relationship between helmet use and positive toxicology screenings on outcomes in ATV accident victims. METHODS: This is a retrospective study of patients admitted to a Level 1 Trauma Center in southwestern West Virginia following an ATV accident between 2005 and 2013. Data were obtained from the institution's Trauma Registry. RESULTS: A total of 1,857 patients were admitted during the study period with 39 (1.9%) reported deaths. Positive serum alcohol and/or urine drug screens were obtained in 66.4% of the patients tested (n=1,293). Those with positive screenings were 9.5% less likely to utilize a helmet (13.2% vs. 22.7%, p<0.001); and the lack of helmet use was associated with an increase in traumatic brain injury (57.1% vs. 41.7%, p<0.001). Positivity for substances or the lack of helmet use was significantly associated with higher morbidity. Lack of helmet use resulted in a 3.94-fold increase in the risk of discharge in a vegetative state or death. CONCLUSIONS: Drugs and alcohol use may predispose riders to be less likely to wear helmets and significantly increase the risk of a poor clinical outcome following an ATV accident. Rigorous efforts should be made to enhance safety measures through educational endeavors and amendment of current regulations to promote safe and responsible use of ATVs. PRACTICAL APPLICATIONS: Modification of regulatory requirements should be considered in order to mandate the wearing of helmets during ATV operation. In addition, expansion of safety programs should be considered in an effort to improve availability, affordability and awareness of safe ATV practices.

INFLUENCE OF LOCALIZER AND SCAN DIRECTION ON THE DOSE-REDUCING EFFECT OF AUTOMATIC TUBE CURRENT MODULATION IN COMPUTED TOMOGRAPHY.

The purpose of this study was to investigate the influence of the localizer and scan direction on the dose-reducing efficacy of the automatic tube current modulation (ATCM) in computed tomography (CT). Craniocaudal and caudocranial chest CT scans, based on anterior-posterior (AP), posterior-anterior (PA), lateral (LAT) or dual AP/LAT localizers, of an anthropomorphic phantom containing thermoluminescent dosimeters (TLDs), were made on three Siemens systems. TLD readings were converted to lung and thyroid doses. A second dose estimation was performed based on Monte Carlo simulations. In addition, the ATCM behaviour of GE and Toshiba was evaluated based on AP, PA and LAT localizers. Compared with AP, tube currents of PA and AP/LAT scans were on average 20 % higher and 40 % lower, respectively, for the Siemens systems. Consequently, thyroid and lung doses increased with 60 % with a PA instead of an AP/LAT scan, with significant differences in image noise. Moreover, the thyroid dose halves by taking the scan in caudocranial direction. Noise values were not significantly different when changing scan direction.