Dual-Energy Computed Tomography Virtual Noncalcium Imaging of Intracranial Arteries in Acute Ischemic Stroke: Differentiation Between Acute Thrombus and Calcification.

OBJECTIVE: Hyperdense artery sign (HAS) on noncontrast brain computed tomography (CT) indicates an acute thrombus within the cerebral artery. It is a valuable imaging biomarker for diagnosing large-vessel occlusion; however, its identification may be challenging with the presence of vascular calcification. Dual-energy CT virtual noncalcium (VNCa) imaging using a 3-material decomposition algorithm is helpful for differentiating between calcification and hemorrhage. This study aimed to clarify the potential of VNCa imaging for differentiating HAS from vascular calcification. METHODS: Patients with acute ischemic stroke and large-vessel occlusion identified on MR angiography, who also underwent noncontrast dual-energy CT, were included. The 80 kV/Sn 140 kV mixed images, with a weighting factor of 0.4, were considered 120 kVp-equivalent images. Postprocessing using a 3-material decomposition algorithm to differentiate between calcium (Ca), cerebrospinal fluid, and hemorrhage was performed via a commercially available 3-dimensional workstation. A mixed image, VNCa image, color-coded Ca image, and color-coded Ca image with VNCa image overlay (color-coded Ca-overlay image) were obtained, and axial reconstruction with a 1-mm slice thickness was performed for each image type. Two experienced neuroradiologists conducted imaging evaluations in consensus. RESULTS: Thirty-four patients (mean age, 76.0 years; 21 male and 13 female patients) were included. The mixed and VNCa images revealed an HAS (indicating an acute clot) corresponding to the large-vessel occlusion site in 30 patients. Among them, the VNCa and color-coded Ca-overlay images enabled clear differentiation between the acute thrombus and adjacent vessel wall calcification in 5 patients. Among the other 4 patients, the VNCa, Ca-overlay, and Ca images identified calcified cerebral emboli in the M1 segment in 1 patient. For the other 3 patients, no high attenuation corresponding to magnetic resonance angiography findings was observed in any of the mixed, VNCa, Ca-overlay, or Ca images. CONCLUSIONS: VNCa and color-coded Ca-overlay images obtained via dual-energy brain CT enabled differentiation of acute thrombus from vessel wall calcification and calcified cerebral emboli in patients with acute ischemic stroke.

Development of an automatic multiplanar reconstruction processing method for head computed tomography.

BACKGROUND: Head computed tomography (CT) is a commonly used imaging modality in radiology facilities. Since multiplanar reconstruction (MPR) processing can produce different results depending on the medical staff in charge, there is a possibility that the antemortem and postmortem images of the same person could be assessed and identified differently. OBJECTIVE: To propose and test a new automatic MPR method in order to address and overcome this limitation. METHODS: Head CT images of 108 cases are used. We employ the standardized transformation of statistical parametric mapping 8. The affine transformation parameters are obtained by standardizing the captured CT images. Automatic MPR processing is performed by using this parameter. The sphenoidal sinus of the orbitomeatal cross section of the automatic MPR processing of this study and the conventional manual MPR processing are cropped with a matrix size of 128×128, and the value of zero mean normalized correlation coefficient is calculated. RESULTS: The computed zero mean normalized cross-correlation coefficient (Rzncc) of≥0.9, 0.8≤Rzncc < 0.9 and 0.7≤Rzncc < 0.8 are achieved in 105 cases (97.2%), 2 cases (1.9%), and 1 case (0.9%), respectively. The average Rzncc was 0.96±0.03. CONCLUSION: Using the proposed new method in this study, MPR processing with guaranteed accuracy is efficiently achieved.

Staphylococcus aureus-induced endothelial permeability and inflammation are mediated by microtubule destabilization.

Staphylococcus aureus is a major etiological agent of sepsis and induces endothelial cell (EC) barrier dysfunction and inflammation, two major hallmarks of acute lung injury. However, the molecular mechanisms of bacterial pathogen-induced EC barrier disruption are incompletely understood. Here, we investigated the role of microtubules (MT) in the mechanisms of EC barrier compromise caused by heat-killed S. aureus (HKSA). Using a customized monolayer permeability assay in human pulmonary EC and MT fractionation, we observed that HKSA-induced barrier disruption is accompanied by MT destabilization and increased histone deacetylase-6 (HDAC6) activity resulting from elevated reactive oxygen species (ROS) production. Molecular or pharmacological HDAC6 inhibition rescued barrier function in HKSA-challenged vascular endothelium. The HKSA-induced EC permeability was associated with impaired MT-mediated delivery of cytoplasmic linker-associated protein 2 (CLASP2) to the cell periphery, limiting its interaction with adherens junction proteins. HKSA-induced EC barrier dysfunction was also associated with increased Rho GTPase activity via activation of MT-bound Rho-specific guanine nucleotide exchange factor-H1 (GEF-H1) and was abolished by HDAC6 down-regulation. HKSA activated the NF-κB proinflammatory pathway and increased the expression of intercellular and vascular cell adhesion molecules in EC, an effect that was also HDAC6-dependent and mediated, at least in part, by a GEF-H1/Rho-dependent mechanism. Of note, HDAC6 knockout mice or HDAC6 inhibitor-treated WT mice were partially protected from vascular leakage and inflammation caused by both HKSA or methicillin-resistant S. aureus (MRSA). Our results indicate that S. aureus-induced, ROS-dependent up-regulation of HDAC6 activity destabilizes MT and thereby activates the GEF-H1/Rho pathway, increasing both EC permeability and inflammation.

[Novel Perfusion Evaluation Method Using Phase-ratio Image Map in Head 4D-CT].

PURPOSE: CT perfusion (CTP) is a powerful tool for the assessment of cerebrovascular disease. However, CTP maps are significantly different depending on CTP software and algorithm, even when using identical image data. We developed a phase-ratio image map (PI map), which was a novel perfusion map, without using CTP software. The purpose of this study was to investigate the usefulness of the PI map by comparing it with a positron emission tomography (PET) image. METHODS: Twenty patients (16 men, 4 women; mean age: 61.6 years) with unilateral cervical and intracranial steno-occlusive disease underwent CTP. CTP source images were obtained at 1-s intervals of 23 times and 5 intervals using dynamic multiphase imaging. An early-phase image was generated by computing the average of CT images for 5 s in the vicinity of the peak enhancement curve of a normal hemisphere. A delayed-phase image was generated by computing the average of CT images for 5 s immediately after the early phase. The PI map was created by dividing the delayed-phase image by the early-phase image. We investigated the validity of the PI map compared with PET-cerebral blood flow (CBF). Lesion-to-normal ratios between a PET-CBF and the PI map or two conventional CTP-CBFs were observed and compared, and the relative errors were also compared. RESULT: There was a strong correlation between the PET-CBF and the PI map (R=0.82). Correlations between the PET-CBF and two CTP-CBFs were weak (R=0.30) and middle (R=0.62), respectively. The relative error between the PI map and the PET-CBF was within 10% in most cases. CONCLUSION: The PI map was more similar to the PET-CBF on perfusion evaluation, and did not depend on CTP software. The robustness and simplicity of the PI mapping method would be advantageous compared with conventional CTP mapping methods.

Otx2 expression in anterior neuroectoderm and forebrain/midbrain is directed by more than six enhancers.

Otx2 plays essential roles in each site at each step of head development. We previously identified the AN1 enhancer at 91kb 5' upstream for the Otx2 expressions in anterior neuroectoderm (AN) at neural plate stage before E8.5, and the FM1 enhancer at 75kb 5' upstream and the FM2 enhancer at 122kb 3' downstream for the expression in forebrain/midbrain (FM) at brain vesicle stage after E8.5. The present study identified a second AN enhancer (AN2) at 88kb 5' upstream; the AN2 enhancer also recapitulates the endogenous Otx2 expression in choroid plexus, cortical hem and choroidal roof. However, the enhancer mutants indicated the presence of another AN enhancer. The study also identified a third FM enhancer (FM3) at 153kb 5' upstream. Thus, the Otx2 expressions in anterior neuroectoderm and forebrain/midbrain are regulated by more than six enhancers located far from the coding region. The enhancers identified are differentially conserved among vertebrates; none of the AN enhancers has activities in caudal forebrain and midbrain at brain vesicle stage after E8.5, nor do any of the FM enhancers in anterior neuroectoderm at neural plate stage before E8.5.

[Examination of Visual Effect in Low-dose Cerebral CT Perfusion Phantom Image Using Iterative Reconstruction].

CT perfusion (CTP) is obtained cerebrovascular circulation image for assessment of stroke patients; however, at the expense of increased radiation dose by dynamic scan. Iterative reconstruction (IR) method is possible to decrease image noise, it has the potential to reduce radiation dose. The purpose of this study is to assess the visual effect of IR method by using a digital perfusion phantom. The digital perfusion phantom was created by reconstructed filtered back projection (FBP) method and IR method CT images that had five exposure doses. Various exposure dose cerebral blood flow (CBF) images were derived from deconvolution algorithm. Contrast-to-noise ratio (CNR) and visual assessment were compared among the various exposure dose and each reconstructions. Result of low exposure dose with IR method showed, compared with FBP method, high CNR in severe ischemic area, and visual assessment was significantly improvement. IR method is useful for improving image quality of low-dose CTP.

Non-contrast dual-energy CT using X-map for acute ischemic stroke: region-specific comparison with simulated 120-kVp CT and diffusion-weighted MR images.

PURPOSE: X-map is a non-contrast dual-energy CT (DECT) application to identify acute ischemic stroke (AIS). Our aim was to verify region-specific characteristics of early ischemic changes (EIC) on X-map compared with simulated 120-kVp mixed-CT image and DWI. METHODS: Fifty AIS patients who underwent DECT and DWI were enrolled (mean age, 76 years; 34 men, 16 women). All datasets including mixed-CT image, X-map, and DWI were transformed into a standard brain atlas with 11 × 2 ROIs based on the ASPECTS + W system. ROIs with EIC on DWI, mixed-CT image, and X-map were defined as DWI-positive, mixed-CT-positive, and X-map-positive, and those with normal finding were DWI-negative, mixed-CT-negative, and X-map-negative respectively, in visual assessment by two neuroradiologists in consensus. RESULTS: EIC on X-maps were visually relevant to those on the other images: of 221 ROIs with mixed-CT-positive and X-map-positive, 198 (89.6%) were DWI-positive. X-map revealed moderate diagnostic accuracy for AIS compared with DWI in ROC curve analysis (AUC = 0.732). X-map identified EIC in deep white matter more sensitively than mixed-CT image: of 15 ROIs with mixed-CT-negative and X-map-positive in W segments, 14 (93.3%) were DWI-positive. X-map often showed EIC in cortical regions that were not detected on the other images: of 67 ROIs with mixed-CT-negative and X-map-positive in I and M1-M6 segments, 47 (70.1%) were DWI-negative. CONCLUSIONS: X-map is useful to detect EIC, especially in deep white matter, and may also provide additional information in acute ischemic lesions where DWI cannot be detected.

Neural tube defects by NUAK1 and NUAK2 double mutation.

BACKGROUND: NUAK1 and NUAK2, members of the AMP-activated protein kinase family of serine/threonine kinases, are prominently expressed in neuroectoderm, but their functions in neurulation have not been elucidated. RESULTS: NUAK1 and NUAK2 double mutants exhibited exencephaly, facial clefting, and spina bifida. Median hinge point was formed, but dorsolateral hinge point formation was not apparent in cranial neural plate; neither apical constriction nor apico-basal elongation took place efficiently in the double mutants during the 5-10-somite stages. Concomitantly, the apical concentration of phosphorylated myosin light chain 2, F-actin, and cortactin was insignificant, and development of acetylated α-tubulin-positive microtubules was poor. However, the distribution of F-actin, cortactin, Shroom3, Rho, myosin heavy chain IIB, phosphorylated myosin light chain 2, α-tubulin, γ-tubulin, or acetylated α-tubulin was apparently normal in the double mutant neuroepithelia at the 5-somite stage. CONCLUSIONS: NUAK1 and NUAK2 complementarily function in the apical constriction and apico-basal elongation that associate with the dorsolateral hinge point formation in cephalic neural plate during the 5- to 10-somite stages. In the double mutant neural plate, phosphorylated myosin light chain 2, F-actin, and cortactin did not concentrate efficiently in apical surfaces, and acetylated α-tubulin-positive microtubules did not develop significantly.

[Reduction of radiation exposure during computed tomography perfusion using an iterative reconstruction method].

The purpose of this study was to evaluate the image noise reduction effect of iterative reconstruction (IR) when used to reduce radiation exposure during computed tomography (CT) perfusion. We scanned a contrast phantom using various radiation doses. Image reconstruction was via filtered back projection (FBP) and IR (adaptive iterative dose reduction 3D: AIDR3D). AIDR3D provided four levels of noise reduction (weak, mild, standard, and strong). We examined the accuracy of CTP map (cerebral blood volume: CBV, mean transist time: MTT, cerebral blood flow: CBF) low-dose IR images to create a digital perfusion phantom that simulates the dynamic curve of ischemic cases using reconstructed images. The optimal filter type of IR was evaluated in the low-frequency area of the NPS at low doses. We were able to obtain the optimal filter type of IR in the low-frequency area of the NPS that was equivalent to that of the reference (150 mA, FBP). The CTP map created using the optimal filter type of IR allowed dose reduction to 80 mA, much lower than the reference. We conclude that it is possible to reduce the dose to 46% of the reference level by using the NPS for dose reduction and IR. IR thus has the potential to contribute to reduction of radiation exposure during CT perfusion.

[A Review of Current Knowledge for X-ray Energy in CT: Practical Guide for CT Technologist].

In computed tomography (CT) systems, the optimal X-ray energy in imaging depends on the material composition and the subject size. Among the parameters related to the X-ray energy, we can arbitrarily change only the tube voltage. For years, the tube voltage has often been set at 120 kVp. However, since about 2000, there has been an increasing interest in reducing radiation dose, and it has led to the publication of various reports on low tube voltage. Furthermore, with the spread of dual-energy CT, virtual monochromatic X-ray images are widely used since the contrast can be adjusted by selecting the optional energy. Therefore, because of the renewed interest in X-ray energy in CT imaging, the issue of energy and imaging needs to be summarized. In this article, we describe the basics of physical characteristics of X-ray attenuation with materials and its influence on the process of CT imaging. Moreover, the relationship between X-ray energy and CT imaging is discussed for clinical applications.

Evolutionary origin of the Otx2 enhancer for its expression in visceral endoderm.

In the mouse, the Otx2 gene has been shown to play essential roles in the visceral endoderm during anterior-posterior axis formation and head induction. While these are primary processes in vertebrate embryogenesis, the visceral endoderm is a tissue unique to mammals. Two enhancers (VE and CM) have been previously found to direct Otx2 expression during early embryogenesis. This study demonstrates that in anterior visceral endoderm the CM enhancer does not have an activity by itself, but enhances the activity of the VE enhancer. These two enhancers also cooperate for the activities in anterior mesendoderm and cephalic mesenchyme. Comparative studies suggest that VE enhancer function was most likely established before the divergence of sarcopterygians into Actinistia, Dipnoi and tetrapods, while the nucleotide sequence corresponding to the VE enhancer was already present in the last common ancestor of bony fishes. The CM enhancer sequence and function would have been also established in ancestral sarcopterygians. The VE/CM enhancers and their gene cascades in the ancestral sarcopterygian head organizer would then have been co-opted by amphibian deep endoderm cells and mammalian visceral endoderm cells for the head development.

Appropriate iMAR presets for metal artifact reduction from surgical clips and titanium burr hole covers on postoperative non-contrast brain CT.

PURPOSE: To evaluate suitable iterative metal artifact reduction (iMAR) presets for titanium neurosurgical clips and burr hole covers (BHCs) on postoperative non-contrast computed tomography (NCCT). METHOD: Twenty-two patients who underwent NCCT after intracranial aneurysmal clipping were included. NCCT images were postprocessed using eight currently available iMAR presets. In each image, a circular region of interest (ROI) was placed around clip, BHC, and on parietal lobe as reference. Standard deviation (SD) and attenuation value (HU) were measured in each ROI to obtain artifact index (AI) and contrast-to-noise ratio (CNR). For each iMAR preset, SD, AI, HU, and CNR were compared with those without iMAR for clips and BHCs. Visual assessment around each clip and BHC was performed by two neuroradiologists using three-point visual score (VS) (1 = no apparent, 2 = minor, and 3 = severe artifacts). RESULTS: Among the presets, the neuro-coils preset (iMAR-NC) showed the lowest SD, AI, and VS for clips (P < 0.001). For BHCs, HU, CNR, and VS with iMAR-NC were significantly higher than those without iMAR (P < 0.001). SD, AI, and VS with the shoulder implants preset (iMAR-ShI) were significantly lower than those without iMAR for clips (P = 0.002, 0.002, and P <  0.001, respectively). For BHCs, VS with iMAR-ShI was lowest among the presets (P = 0.004). CONCLUSIONS: Although iMAR-NC reduces metal artifacts from clips, it strengthens artifacts from BHCs. For postoperative NCCT, iMAR-ShI most effectively reduces metal artifacts from both clips and BHCs in a single preset.

Whole-brain perfusion measurement using 320-detector row computed tomography in patients with cerebrovascular steno-occlusive disease: comparison with 15O-positron emission tomography.

OBJECTIVE: The 320-detector row computed tomography (CT) can provide whole-brain CT perfusion (CTP) maps with continuous angiographic images by performing a single dynamic scan. We investigated the reliability of CTP cerebral blood flow (CTP-CBF) with 320-detector row CT by comparing findings with O-positron emission tomography (PET-CBF). METHODS: Whole-brain CTP and PET were performed in 10 patients with chronic unilateral steno-occlusive disease. We compared absolute and relative CBF values of bilateral middle cerebral artery territories between CTP and PET. RESULTS: Although mean CTP-CBF values were approximately 30% lower than mean PET-CBF values, the mean ischemic-to-nonischemic CBF ratios of CTP and PET were almost identical (P = 0.804). Regression analysis showed a significant correlation between CTP-CBF and PET-CBF values for each patient (r = 0.52-0.85, P < 0.001). CONCLUSIONS: Whole-brain CTP using 320-detector row CT is useful for evaluating the degree of ischemia for the entire brain with chronic cerebrovascular disease.

Development of a deep learning model to identify hyperdense MCA sign in patients with acute ischemic stroke.

PURPOSE: The aim of this study was to develop an interactive deep learning-assisted identification of the hyperdense middle cerebral artery (MCA) sign (HMCAS) on non-contrast computed tomography (CT) among patients with acute ischemic stroke. MATERIALS AND METHODS: 35 HMCAS-positive and 39 HMCAS-negative samples extracted by 50-pixel-diameter circular regions of interest were obtained as training and validation datasets according to the consensus decisions of two experienced neuroradiologists. Data augmentation was performed to increase the number of training samples. A deep convolutional neural network (DCNN) (Xception) was used to classify input images as HMCAS-positive or -negative. Leave-one-case-out cross-validation was achieved to estimate sensitivity, specificity, and accuracy of the deep learning-based training model for identifying HMCAS. RESULTS: In terms of diagnostic performance, DCNN for HMCAS offered 82.9% sensitivity, 89.7% specificity, and 86.5% accuracy in leave-one-case-out cross-validation. Area under the receiver operating characteristic curve for HMCAS was 0.947 (95% confidence interval 0.895-0.998; P < 0.05). CONCLUSION: The deep learning method appears potentially beneficial for identifying HMCAS on non-contrast CT in patients with acute ischemic stroke.

Evaluation of lumbar intervertebral disc degeneration using dual energy CT virtual non-calcium imaging.

PURPOSE: To clarify the utility of dual energy CT (DECT) virtual non-calcium (VNCa) imaging for investigating lumbar intervertebral disc degeneration. METHOD: Fifty-three patients who underwent both DECT and MR imaging were retrospectively reviewed. Midsagittal T2-weighted imaging findings of all discs were classified based on modified Pfirrmann grade (mPG). Quantitative evaluation of VNCa maps was achieved by setting volumes of interest on each disc. We compared VNCa CT values with mPG using Spearman's rank correlation and one-way ANOVA. VNCa imaging findings of each disc were classified by two neuroradiologists into one of three categories based on the attenuation of nucleus pulposus (NP) compared to that of annulus fibrosus (AF) or muscle. The relationship between the visual categories for each rater and mPGs was analyzed by chi-square test. Statistical significance was established at P < 0.05. RESULTS: Among the included 171 lumbar discs, significant positive correlation was found between VNCa CT values for NP and mPGs (R2 = 0.574, P < 0.05), whereas no significant correlation was found between those for AF and mPGs (R2= -0.015, P = 0.846). Mean VNCa CT values for NP were significantly different among each mPG (P < 0.05 for each), except between grades 3 and 4 (P = 0.111). Mean VNCa CT values for AF were not significantly different among each mPG (P = 0.160-1.000). Statistically significant difference was observed among the visual categories for VNCa maps and mPGs in each rater (P < 0.05 for both). CONCLUSIONS: VNCa imaging acquired by a single DECT scan and post-processing has potential as an imaging biomarker of lumbar intervertebral disc degeneration.

Usefulness of deep learning-assisted identification of hyperdense MCA sign in acute ischemic stroke: comparison with readers' performance.

PURPOSE: To evaluate the usefulness of deep learning-assisted diagnosis for identifying hyperdense middle cerebral artery sign (HMCAS) on non-contrast computed tomography in comparison with the diagnostic performance of neuroradiologists. MATERIALS AND METHODS: We obtained 46 HMCAS-positive and 52 HMCAS-negative test samples extracted using 50-pixel-diameter circular regions of interest. Five neuroradiologists undertook an initial diagnostic performance test by describing the HMCAS-positive prediction rate in each sample. Their diagnostic performance was compared with that of a deep convolutional neural network (DCNN) model that had been trained using another dataset in our previous study. In the second test, readers could reference the prediction rate of the DCNN model in each sample. RESULTS: The diagnostic performance of the DCNN for HMCAS showed an accuracy of 81.6% and area under the receiver-operating characteristic curve (AUC) of 0.869, whereas the initial diagnostic performance of neuroradiologists showed an accuracy of 78.8% and AUC of 0.882. The second diagnostic test of neuroradiologists with reference to the results of the DCNN model showed an accuracy of 84.7% and AUC of 0.932. In all readers, AUC values were higher in the second test than the initial test. CONCLUSION: The ability of DCNN to identify HMCAS is comparable with the diagnostic performance of neuroradiologists.

MicroRNA-9 modulates Cajal-Retzius cell differentiation by suppressing Foxg1 expression in mouse medial pallium.

Vertebrate brain hosts a diverse collection of microRNAs, but little is known about their functions in vivo. Here we propose that mouse microRNA-9 (miR-9) targets Foxg1 mRNAs for proper generation of Cajal-Retzius cells in the medial pallium. miR-9 expression is mediolaterally graded, being most intense in the cortical hem; it contrasts with the Foxg1 expression in a reciprocal gradient. The 3' untranslated regions of tetrapod, but not of teleost, Foxg1 mRNAs conserve miR-9 target sequences and are regulated by miR-9. Gain- and loss-of-function analyses of miR-9 showed that miR-9 negatively regulates endogenous Foxg1 protein level. Moreover, miR-9 overexpression in developing telencephalon at E11.5 by electroporation resulted in ectopic Reelin-positive cells over the cortex beyond the marginal zone. In addition, inhibition of endogenous miR-9 function by antisense oligonucleotides caused the regression of Wnt3a-positive cortical hem and reduction of reelin-, p73-, and NeuroD1-positive cells.

Microtubule destabilization caused by particulate matter contributes to lung endothelial barrier dysfunction and inflammation.

Exposure to particulate matter (PM) associated with air pollution remains a major public health concern, as it has been linked to significant increase in cardiopulmonary morbidity and mortality. Lung endothelial cell (EC) dysfunction is one of the hallmarks of cardiovascular events of lung exposure to PM. However, the role of PM in acute lung injury (ALI) exacerbation and delayed recovery remains incompletely understood. This study tested a hypothesis that PM augments lung injury and EC barrier dysfunction via microtubule-dependent mechanisms. Our data demonstrate that in pulmonary EC PM caused time- and dose-dependent remodeling of actin cytoskeleton and considerable destabilization of the microtubule (MT) network. These events led to the weakening of cell junctions and formation of actin stress fibers, resulting in disruption of lung EC monolayer and increased permeability. PM also caused ROS-dependent activation of MT-specific deacetylase, HDAC6. Suppression of HDAC6 activity by pharmacological inhibitors or siRNA-based depletion of HDAC6 abolished PM-induced EC permeability increase, which was accompanied by reduced activation of stress kinase signaling, inhibition of Rho cascade, decreased IL-6 production and suppressed activation of its downstream target STAT3. Pretreatment of pulmonary EC with IL-6 inhibitor led to inhibition of STAT3 activity and decreased PM-induced hyper-permeability. Because one of the major activators of Rho-GTPase, GEFH1, is localized on the MT, we examined its involvement in PM-caused EC barrier compromise. Inhibition of GEF-H1 activation significantly attenuated PM-induced permeability increase. Moreover, combined inhibition of IL-6 and GEF-H1 signaling exhibited additive protective effect. Taken together, these results demonstrate a critical involvement of MT-associated signaling in the PM-induced exacerbation of lung EC barrier compromise and inflammatory response.

Role of truncated oxidized phospholipids in acute endothelial barrier dysfunction caused by particulate matter.

Particulate matter (PM) air pollution is a global environmental health problem contributing to more severe lung inflammation and injury. However, the molecular and cellular mechanisms of PM-induced exacerbation of lung barrier dysfunction and injury are not well understood. In the current study, we tested a hypothesis that PM exacerbates vascular barrier dysfunction via ROS-induced generation of truncated oxidized phospholipids (Tr-OxPLs). Treatment of human pulmonary endothelial cells with PM caused endothelial cell barrier disruption in a dose-dependent fashion. Biochemical analysis showed destabilization of cell junctions by PM via tyrosine phosphorylation and internalization of VE-cadherin. These events were accompanied by PM-induced generation of Tr-OxPLs, detected by mass spectrometry analysis. Furthermore, purified Tr-OxPLs: POVPC, PGPC and lyso-PC alone, caused a rapid increase in endothelial permeability and augmented pulmonary endothelial barrier dysfunction induced by submaximal doses of PM. In support of a role of TR-OxPLs-dependent mechanism in mediation of PM effects, ectopic expression of intracellular type 2 platelet-activating factor acetylhydrolase (PAFAH2), which specifically hydrolyzes Tr-OxPLs, significantly attenuated PM-induced endothelial hyperpermeability. In summary, this study uncovered a novel mechanism of PM-induced sustained dysfunction of pulmonary endothelial cell barrier which is driven by PM-induced generation of truncated products of phospholipid oxidation causing destabilization of cell junctions.