Lung Blood Vessels Research Articles

Mechanosensitive ATP release in the lungs: New insights from real-time luminescence imaging studies.

Extracellular ATP and other nucleotides are important autocrine/paracrine mediators that stimulate purinergic receptors and regulate diverse processes in the normal lungs. They are also associated with pathogenesis of a number of respiratory diseases and clinical complications including acute respiratory distress syndrome and ventilator induced lung injury. Mechanical forces are major stimuli for cellular ATP release but precise mechanisms responsible for this release are still debated. The present review intends to provide the current state of knowledge of the mechanisms of ATP release in the lung. Putative pathways of the release, including the contribution of cell membrane injury and cell lysis are discussed addressing their strength, weaknesses and missing evidence that requires future study. We also provide an overview of the recent technical advances in studying cellular ATP release in vitro and ex vivo. Special attention is given to new insights into lung ATP release obtained with the real-time luminescence ATP imaging. This includes recent data on stretch-induced mechanosensitive ATP release in a model and primary cells of lung alveoli in vitro as well as inflation-induced ATP release in airspaces and pulmonary blood vessels of lungs, ex vivo.

Abstract 17091: High Efficient Therapeutic Genome Editing in Cardiovascular Endotheliumby Nanoparticle Delivery of Crispr Plasmid

Introduction: Therapeutic delivery of CRISPR system components to induce in vivo genome editing in postnatal life has great translational potential. Recent studies employing non-viral delivery of small guide RNA (gRNA) and Cas9 mRNA have achieved efficient in vivo genome editing in adult mice. However, as often seen in other RNA therapeutic studies with non-viral delivery of antisense and siRNA, the efficiency is limited to the liver. Hypothesis: Novel nanoparticle can deliver CRISPR system components in vivo to selectively target cardiovascular endothelium in adult mice. Methods: We developed a novel nanoparticles. Mixture of the nanoparticle:plasmid DNA expressing Cas9 under the control of the human CDH5 promoter and gRNA driven by the U6 promoter was administered i.v. to adult mice. Seven to ten days later, various organ tissues were collected for analysis of the efficiency of genomic editing and knockout of protein expression. The phenotype of CRISPR-mediated in vivo knockout of Pik3cg which encodes the G protein-coupled receptor-activated p110gamma isoform of PI3K was compared to Pik3cg null mice in response to sepsis challenge. Results: Therapeutic delivery of nanoparticles loaded with the all-in-one CRISPR plasmid DNA induced highly efficient genome editing in endothelial cells (ECs) of the cardiovascular system including lung, heart, aorta, and peripheral blood vessels in adult mice. The Indel rate was as great as 50% in ECs isolated from these vascular beds. Immunostaining and Western blotting demonstrated greater than 70% decrease of protein expression in cardiovascular endothelial cells. Pik3cg -gRNA-induced genome editing diminished p110γPI3K expression in pulmonary vascular ECs, which led to impaired vascular repair and resolution of inflammation after sepsis challenge as seen in Pik3cg -/- mice. Conclusion: We have developed a simple and highly efficient method for in vivo genome editing selectively in the cardiovascular endothelium. This strategy will greatly facilitate cardiovascular research and may enable therapeutic genome editing for prevention and treatment of cardiovascular diseases.

Rapamycin alleviated pulmonary injury induced by fat embolism syndrome in rats

Fat embolism syndrome (FES) is a serious complication after trauma, surgery and fat emulsion input and can lead to serious pulmonary injury. Autophagy controls the cell survival and homeostasis by removing the mis-folded proteins and damaged organelles as well as intracellular pathogens through a lysosomal degradation pathway. Increasing research documented that autophagy was wildly involved in variety of human diseases and had huge therapeutic potential. However, the role and mechanism of autophagy in FES remains largely unknown. The rat model of FES was established by tail vein injection with fat and was assessed by Wet-to-Dry (W/D) ratio analysis, hematoxylin-eosin (HE) analysis, staining Oil red staining analysis and qPCR analysis. Western blots were employed to detect the expression of autophagy markers. The changes of pulmonary injury were observed after premedication of rapamycin (an autophagy activator). The alveolar structural damage, red free fat substances in the blood vessels of lung, increased the lung ratio, and the up-regulated MPO expression and activity were showed in the FES models. The expressions of autophagy markers were decreased and meanwhile, apoptosis markers were increased in the FES model. Rapamycin restored the expression of autophagy markers and inhibited the apoptosis and further, resulting in the improvement of the pulmonary injury. Thus, our study demonstrated that autophagy was inhibited and apoptosis was promoted in FES and further Rapamycin alleviated the pulmonary damage in FES via restoring the autophagy and inhibiting the apoptosis.

Features of reorganization of blood vessels of lungs of rats at various degrees of total dehydration.

Актуальність. Вода з розчиненими в ній речовинами є функціонально єдиною системою організму, яка забезпечує реалізацію гомеостазу. Постійний рівень осмотичного тиску плазми крові, міжклітинної і внутрішньоклітинної рідини є однією з головних умов нормальної життєдіяльності організму. Порушення гідроелектролітної константи відбуваються в організмі майже постійно, але іноді вони виходять за рамки середньостатистичних норм. Одним з таких проявів є дефіцит води в організмі, що виникає внаслідок найрізноманітніших фізіологічних та патологічних станів. Безпосереднім наслідком зневоднення буває порушення периферійного кровотоку з розвитком циркуляторної гіпоксії. Мета дослідження: встановити особливості реорганізації кровоносних судин легень щурів за різних ступенів загального зневоднення та дати їм кількісну характеристику. Методи. Експерименти проведено на білих лабораторних статевозрілих щурах-самцях яким моделювали загальну дегідратацію легкого, середнього і важкого ступеня. Проводили гістологічне і морфометричне дослідження кровоносних судин легень. Результати. У першу добу експерименту спостерігалося посилення кровопостачання легень, яке підтверджувалося тенденцією до розширення просвітів як легеневих, так і бронхіальних артерій, що підтверджувалося зниженням у них рівня функціонального показника – індексу Вогенворта. Проте у наступні терміни спостерігалася висхідна вазоконстрикція з одночасним зниженням пропускної здатності артерій дрібного і середнього калібру та розширенням просвіту і збільшенням ємності артерій великого калібру. Вени, навпаки, поступово ставали малокровними, що супроводжувалося підвищенням тонусу їх стінки і звуженням просвіту, який нерідко набував «зірчастої» деформації. Із збільшенням терміну спостереження і відповідно ступеня важкості загального зневоднення збільшувалася частота виявлення артерій «замикаючого» типу. Підсумок. Результати проведеного дослідження свідчать, що при загальному зневодненні організму поступово у паренхімі легень і їх кровоносних руслах розвиваються певні морфофункціональні зміни гіповолемічного характеру інтенсивність яких залежить від тривалості зневоднення, а значить його ступеня.

Discrete Heat Kernel Smoothing in Irregular Image Domains.

We present the discrete version of heat kernel smoothing on graph data structure. The method is used to smooth data in an irregularly shaped domains in 3D images. New statistical properties of heat kernel smoothing are derived. As an application, we show how to filter out noisy data in the lung blood vessel trees obtained from computed tomography. The method can be further used in representing the complex vessel trees parametrically as a linear combination of basis functions and extracting the skeleton representation of the trees.

СТРЕССОРНОЕ ЛЕГКОЕ КРЫС КАК ЭКСПЕРИМЕНТАЛЬНАЯ МОДЕЛЬ ЛЕГОЧНОЙ ГИПЕРТЕНЗИИ И ГИПЕРЕМИИ

A review of the literature is devoted to the peculiarities of the lung structure of white rats. Morphological features of rat lungs are an experimental model in the case of the study of the development of left ventricular pulmonary hypertension. Rats have the features of the lungs structure, which differ from the lungs of man. The construction of blood vessels of the rat lungs should be studied in the case of a stressor lung. Peculiarities of innervation of the pulmonary veins of rats in the root of the lung make the left lung of rats an object of study necessary for solving the problems of modern pulmonology, cardiology and morphology. In the anatomical study, the root of the diaphragmatic lobe is located in the caudal sulcus of the left lung. In studying the features of the sintopia and holotopia of the root and gates of the left lung of sexually mature rats, the structural apparatus of the caudal sulcus is discovered, which is located for more than 11-17 mm on the medial surface of the diaphragmatic lobe of the left lung. The structural apparatus consists of the left caudal pulmonary vein, furrows on the surface of the diaphragmatic lobe of the left lung, the adventitial shell of the caudal bronchus, the visceral pleura, the bronchial nerves and blood vessels, the encapsulated receptors. The presence of cardiomyocytes in the intrapulmonary veins in rats confirms the hypothesis of a rhythmic, valve-like action of the transverse striated muscle of the pulmonary venous wall during systole and a possible role in pulmonary circulation. Data obtained through experimental intervention indicate the valve-like effect of the striated muscle of the pulmonary venous wall.

Regulation of eosinophil recruitment and allergic airway inflammation by heparan sulfate proteoglycan (HSPG) modifying enzymes

ABSTRACTBackground: HSPGs are glycoproteins containing covalently attached heparan sulfate (HS) chains which bind to growth factors, chemokines, etc., and regulate various aspects of inflammation including cell recruitment. We previously showed that deletion of endothelial N-acetylglucosamine N-deacetylase-N-sulfotransferase-1 (Ndst1), an enzyme responsible for N-sulfation during HS biosynthesis, reduces allergic airway inflammation (AAI). Here, we investigated the importance of O-sulfation mediated by uronyl 2-O-sulfotransferase (Hs2st) in development of AAI relative to N-sulfation. Methods: Mice deficient in endothelial and leukocyte Hs2st (Hs2stf/fTie2Cre+) or Ndst1 (Ndst1f/fTie2Cre+) and WT mice were challenged with Alternaria alternata and evaluated for airway inflammation. Trafficking of murine eosinophils on lung endothelial cells was examined in vitro under conditions of flow. Results: Exposure to Alternaria decreased expression level of Hs2st in WT mice while level of Ndst1 remained unchanged. Compared to WT mice, Alternaria-challenged Hs2stf/fTie2Cre+ mice exhibited significantly increased eosinophils in the bone marrow, bronchoalveolar lavage fluid [BALF] and lung tissue associated with persistent airway hyperresponsiveness, airway mucus hypersecretion and elevated Th2 cytokines. In contrast, Alternaria-challenged Ndst1f/fTie2Cre+ mice exhibited a marked reduction in airway eosinophilia, mucus secretion and smooth muscle mass compared to WT counterparts. While BALF eotaxins were lower in Alternaria-challenged Hs2stf/fTie2Cre+ relative to WT mice, they were not reduced to background levels as in allergen-challenged Ndst1f/fTie2Cre+ mice. Trafficking of murine eosinophils under conditions of flow in vitro was similar on Hs2st-deficient and WT endothelial cells. Expression of ZO-1 in Hs2st-deficient lung blood vessels in control and allergen-challenged mice was significantly lower than in WT counterparts. Conclusions: Our study demonstrates that allergen exposure reduces expression of Hs2st; loss of uronyl 2-O-sulfation in endothelial and leukocyte HSPG amplifies recruitment of eosinophils likely due to a compromised vascular endothelium resulting in persistent inflammation whereas loss of N-sulfation limits eosinophilia and attenuates inflammation underscoring the importance of site-specific sulfation in HSPG to their role in AAI.

Comparison of photon volumetric modulated arc therapy, intensity-modulated proton therapy, and intensity-modulated carbon ion therapy for delivery of hypo-fractionated thoracic radiotherapy

PurposeThe aim of the present study was to compare the dose distribution generated from photon volumetric modulated arc therapy (VMAT), intensity modulated proton therapy (IMPT), and intensity modulated carbon ion therapy (IMCIT) in the delivery of hypo-fractionated thoracic radiotherapy.Methods and materialsTen selected patients who underwent thoracic particle therapy between 2015 and 2016 were re-planned to receive a relative biological effectiveness (RBE) weighted dose of 60 Gy (i.e., GyE) in 15 fractions delivered with VMAT, IMPT, or IMCIT with the same optimization criteria. Treatment plans were then compared.ResultsThere were no significant differences in target volume dose coverage or dose conformity, except improved D95 was found with IMCIT compared with VMAT (p = 0.01), and IMCIT was significantly better than IMPT in all target volume dose parameters. Particle therapy led to more prominent lung sparing at low doses, and this result was most prominent with IMCIT (p < 0.05). Improved sparing of other thoracic organs at risk (OARs) was observed with particle therapy, and IMCIT further lowered the D1cc and D5cc for major blood vessels, as compared with IMPT (p = 0.01).ConclusionAlthough it was comparable to VMAT, IMCIT led to significantly better tumor target dose coverage and conformity than did IMPT. Particle therapy, compared with VMAT, improved thoracic OAR sparing. IMCIT, compared with IMPT, may further improve normal lung and major blood vessel sparing under limited respiratory motion.

ВПЛИВ КСЕНОБІОТИКІВ У ПОВІТРІ НА ПЕРЕБІГ АНГІОПАТІЇ У ЩУРІВ З МОДЕЛЛЮ СИСТЕМНОГО АУТОІМУННОГО РЕВМАТИЧНОГО ЗАХВОРЮВАННЯ

Системні аутоімунні ревматичні захворювання (САРЗ) пов'язані з несприятливими факторами навколишнього середовища, які визначають темпи прогресування і виживання хворих. Забруднення атмосфери полютантами сприяє збільшенню в таких регіонах поширеності цих хвороб.Мета дослідження – оцінити вплив ксенобіотиків у повітрі на характер морфологічних змін судин легень, серця і нирок у тварин з моделлю САРЗ. Матеріали та методи. Експеримент проводили на безпородних щурах, яких поділили на три групи: в першу увійшли тварини з моделлю ССД, які перебували щодня по п'ять годин у несприятливому атмосферному середовищі (аміак+бензол+формалін), другу склали тварини з моделлю захворювання, які знаходилися в звичайних екологічних умовах, а в третю увійшли інтактні щури, яких поміщали в середовище з високим вмістом у повітрі ксенобіотиків, як і представників першої групи. Гістологічні зрізи тканин серця, легень і нирок фарбували гематоксиліном–еозином, альциановим синім і за ван-Гізон, ставили PAS-реакцію.Результати. Зміни з боку легень, серця і нирок виявлені у всіх тварин з експериментальним САРЗ і, відповідно, у 47 %, 47 % і 40 % спостережень інтактних щурів, які перебувають в несприятливому середовищі з забрудненням повітря ксенобіотиками. Негативна екологічна ситуація підвищувала частоту таких морфологічних ознак захворювання як проліферація ендотелію судин серця на 68 % і ниркових артеріол на 52 %, до того ж, існували прямі кореляційні зв'язки між ступенем ангіопатії в окремих органах, що залежало від характеру моделювання патологічного процесу і доводило, що забруднення атмосфери є фактором ризику захворювання у людей. Безпосередньо з моделлю САРЗ був пов'язаний вплив склерозу легеневих судин на розвиток бронхосклерозу, периваскулярної інфільтрації м'яза серця, тоді як забруднення повітря ксенобіотиками визначало залежність ступеня клітинної інфільтрації альвеолярних перегородок від периваскулярною легеневої інфільтрації, розвиток гіпертрофії кардіоміоцитів – від проліферації ендотелію судин серця, збільшення мезангіального матриксу нирок від проліферації ендотелію клубочковох капілярів.Висновок. Забруднення атмосфери ксенобіотиками слід розглядати як негативний фактор щодо розвитку і подальшого перебігу САРЗ у людей, а в патогенезі його провідне значення має ангіопатія.

Murine systemic thrombophilia and hemolytic uremic syndrome from a factor H point mutation

AbstractComplement plays a key role in host defense, but its dysregulation can cause autologous tissue injury. Complement activation is normally controlled by regulatory proteins, including factor H (FH) in plasma and membrane cofactor protein (MCP) on the cell surface. Mutations in FH and MCP are linked to atypical hemolytic uremic syndrome, a type of thrombotic microangiopathy (TMA) that causes renal failure. We describe here that disruption of FH function on the cell surface can also lead to disseminated complement-dependent macrovascular thrombosis. By gene targeting, we introduced a point mutation (W1206R) into murine FH that impaired its interaction with host cells but did not affect its plasma complement-regulating activity. Homozygous mutant mice carrying this mutation developed renal TMA as well as systemic thrombophilia involving large blood vessels in multiple organs, including liver, lung, spleen, and kidney. Approximately 30% of mutant mice displayed symptoms of stroke and ischemic retinopathy, and 48% died prematurely. Genetic deficiency of complement C3 and factor D prevented both the systemic thrombophilia and renal TMA phenotypes. These results demonstrate a causal relationship between complement dysregulation and systemic angiopathy and suggest that complement activation may contribute to various human thrombotic disorders involving both the micro- and macrovasculature.

Histopathological and diagnostic study of Toxoplasmosis in human and sheep by using ELISA in Kut city

This study is designed to diagnosis blood Toxoplasmosis in human (women and men) and sheep (male and female) by using Elisa test; as well histopathological examination of heart, liver and lung of sheep were also done, internal organs were also obtained heart, liver and lung. Women diagnosis was performed by collection 96 samples from aborted women ageing from 15-45 years and divided into three groups; first group 15-25 years involved 50 sample, second group 25-35 years include 37 samples; and the third group 35-45 years included 9 samples. The results of Elisa test showed six positive of IgG, (two for each group) and one positive IgM for the second group, a significant (P&lt;0.05) difference in the first and third groups between IgG and IgM was found. The total percentage of the infection 4, 8.1and 22.2 for the first, second and third groups respectively, and Total of 96 blood samples were collected from men with different chronic diseases and were divided into four groups, (first, second and third) each containing 25 samples and suffering from Thalasemia, renal failure and cancer respectively; of them 21 samples collected from normal men as fourth control group. The results of Elisa diagnostic test indicated 9, 9, 7 and 2 positive of IgG for first, second, third and fourth groups respectively, and 1, 2, 0 and 1 positive of IgM. The total percentage of Toxoplasmosis infection 38.4, 44, 28 and 14.28 for the first, second, third and fourth groups respectively, with a difference between them except the second group appeared significantly P(0.05). 92 samples of sheep blood were collected from male 46 and 46 female. The results of Toxoplasmosis showed one case positive IgG for female and one for male only. The histopathological examination of sheep positive case with chronic infection revealed pathological changes in the organs, the heart showed multiple variable sizes of parasite cysts embedded and scatter between cardiac muscle fibers, in addition to aggregation of inflammatory cells around the blood vessels forming nests of inflammatory cells with congestion of blood vessels in liver and lung. In conclusion that the infection with Toxoplasmosis in Kut city revealed the percentage of infection in men more than the aborted women, also it reported few cases of infection in sheep.

Cytostretch, an Organ-on-Chip Platform.

Organ-on-Chips (OOCs) are micro-fabricated devices which are used to culture cells in order to mimic functional units of human organs. The devices are designed to simulate the physiological environment of tissues in vivo. Cells in some types of OOCs can be stimulated in situ by electrical and/or mechanical actuators. These actuations can mimic physiological conditions in real tissue and may include fluid or air flow, or cyclic stretch and strain as they occur in the lung and heart. These conditions similarly affect cultured cells and may influence their ability to respond appropriately to physiological or pathological stimuli. To date, most focus has been on devices specifically designed to culture just one functional unit of a specific organ: lung alveoli, kidney nephrons or blood vessels, for example. In contrast, the modular Cytostretch membrane platform described here allows OOCs to be customized to different OOC applications. The platform utilizes silicon-based micro-fabrication techniques that allow low-cost, high-volume manufacturing. We describe the platform concept and its modules developed to date. Membrane variants include membranes with (i) through-membrane pores that allow biological signaling molecules to pass between two different tissue compartments; (ii) a stretchable micro-electrode array for electrical monitoring and stimulation; (iii) micro-patterning to promote cell alignment; and (iv) strain gauges to measure changes in substrate stress. This paper presents the fabrication and the proof of functionality for each module of the Cytostretch membrane. The assessment of each additional module demonstrate that a wide range of OOCs can be achieved.

VCAM-1 is a TGF-β1 inducible gene upregulated in idiopathic pulmonary fibrosis

Idiopathic pulmonary fibrosis (IPF) is a chronic lethal interstitial lung disease of unknown etiology. We previously reported that high plasma levels of vascular cell adhesion molecule 1 (VCAM-1) predict mortality in IPF subjects. Here we investigated the cellular origin and potential role of VCAM-1 in regulating primary lung fibroblast behavior. VCAM-1 mRNA was significantly increased in lungs of subjects with IPF compared to lungs from control subjects (p=0.001), and it negatively correlated with two markers of lung function, forced vital capacity (FVC) and pulmonary diffusion capacity for carbon monoxide (DLCO). VCAM-1 protein levels were highly expressed in IPF subjects where it was detected in fibrotic foci and blood vessels of IPF lung. Treatment of human lung fibroblasts with TGF-β1 significantly increased steady-state VCAM1 mRNA and protein levels without affecting VCAM1 mRNA stability. Further, cellular depletion of VCAM-1 inhibited fibroblast cell proliferation and reduced G2/M and S phases of the cell cycle suggestive of cell cycle arrest. These effects on cell cycle progression triggered by VCAM1 depletion were associated with reductions in levels of phosphorylated extracellular regulated kinase 1/2 and cyclin D1. Thus, these observations suggest that VCAM-1 is a TGF-β1 responsive mediator that partakes in fibroblast proliferation in subjects with IPF.

Embolismo tumoral pulmonar microvascular en paciente con tumor urotelial

Pulmonary tumor embolisms (PTE) are an infrequent cause of dyspnea in oncological patients. The majority are diagnosed in patients with advanced tumors, above all localized in the breast, lung or stomach. There are few published cases involving patients with urothelial tumors. We present the case of a 69 year-old male, without a previous diagnosis of cancer, who was admitted due to subacute dyspnea, with clinical suspicion of pulmonary thromboembolism (PT). The patient died on the fifth day of admission. The autopsy confirmed the existence of a tumor in the left renal pelvis with hepatic and lymphoganglionary metastasis and an extensive microvascular pulmonary embolism that affected a large part of the capillaries and medium-caliber blood vessels of both lungs. PTE were considered responsible for the progressive respiratory failure and as the final cause of death. The most frequent clinical presentation of PTE is dyspnea. They are often mistaken for PT and diagnosis is not easy. Their prognosis is very bad, with extremely high mortality and confirmation is usually post-mortem.

Lung dynamic MRI deblurring using low-rank decomposition and dictionary learning.

Lung dynamic MRI (dMRI) has emerged to be an appealing tool to quantify lung motion for both planning and treatment guidance purposes. However, this modality can result in blurry images due to intrinsically low signal-to-noise ratio in the lung and spatial/temporal interpolation. The image blurring could adversely affect the image processing that depends on the availability of fine landmarks. The purpose of this study is to reduce dMRI blurring using image postprocessing. To enhance the image quality and exploit the spatiotemporal continuity of dMRI sequences, a low-rank decomposition and dictionary learning (LDDL) method was employed to deblur lung dMRI and enhance the conspicuity of lung blood vessels. Fifty frames of continuous 2D coronal dMRI frames using a steady state free precession sequence were obtained from five subjects including two healthy volunteer and three lung cancer patients. In LDDL, the lung dMRI was decomposed into sparse and low-rank components. Dictionary learning was employed to estimate the blurring kernel based on the whole image, low-rank or sparse component of the first image in the lung MRI sequence. Deblurring was performed on the whole image sequences using deconvolution based on the estimated blur kernel. The deblurring results were quantified using an automated blood vessel extraction method based on the classification of Hessian matrix filtered images. Accuracy of automated extraction was calculated using manual segmentation of the blood vessels as the ground truth. In the pilot study, LDDL based on the blurring kernel estimated from the sparse component led to performance superior to the other ways of kernel estimation. LDDL consistently improved image contrast and fine feature conspicuity of the original MRI without introducing artifacts. The accuracy of automated blood vessel extraction was on average increased by 16% using manual segmentation as the ground truth. Image blurring in dMRI images can be effectively reduced using a low-rank decomposition and dictionary learning method using kernels estimated by the sparse component.

Concerted activation of the AIM2 and NLRP3 inflammasomes orchestrates host protection against Aspergillus infection.

Invasive pulmonary aspergillosis is a leading cause of infection-associated mortality in immunocompromised individuals. Aspergillus fumigatus infection produces ligands that could activate inflammasomes, but the contribution of these host defenses remains unclear. We show that two inflammasome receptors, AIM2 and NLRP3, recognize intracellular A. fumigatus and collectively induce protective immune responses. Mice lacking both AIM2 and NLRP3 fail to confine Aspergillus hyphae to inflammatory foci, leading to widespread hyphal dissemination to lung blood vessels. These mice succumb to infection more rapidly than WT mice or mice lacking a single inflammasome receptor. AIM2 and NLRP3 activation initiates assembly of a single cytoplasmic inflammasome platform, composed of the adaptor protein ASC along with caspase-1 and caspase-8. Combined actions of caspase-1 and caspase-8 lead to processing of pro-inflammatory cytokines IL-1β and IL-18 that critically control the infection. Thus, AIM2 and NLRP3 form a dual cytoplasmic surveillance system that orchestrates responses against A. fumigatus infection.

Type-1 pericytes accumulate after tissue injury and produce collagen in an organ-dependent manner.

IntroductionFibrosis, or scar formation, is a pathological condition characterized by excessive production and accumulation of collagen, loss of tissue architecture, and organ failure in response to uncontrolled wound healing. Several cellular populations have been implicated, including bone marrow-derived circulating fibrocytes, endothelial cells, resident fibroblasts, epithelial cells, and recently, perivascular cells called pericytes. We previously demonstrated pericyte functional heterogeneity in skeletal muscle. Whether pericyte subtypes are present in other tissues and whether a specific pericyte subset contributes to organ fibrosis are unknown.MethodsHere, we report the presence of two pericyte subtypes, type-1 (Nestin-GFP-/NG2-DsRed+) and type-2 (Nestin-GFP+/NG2-DsRed+), surrounding blood vessels in lungs, kidneys, heart, spinal cord, and brain. Using Nestin-GFP/NG2-DsRed transgenic mice, we induced pulmonary, renal, cardiac, spinal cord, and cortical injuries to investigate the contributions of pericyte subtypes to fibrous tissue formation in vivo.ResultsA fraction of the lung’s collagen-producing cells corresponds to type-1 pericytes and kidney and heart pericytes do not produce collagen in pathological fibrosis. Note that type-1, but not type-2, pericytes increase and accumulate near the fibrotic tissue in all organs analyzed. Surprisingly, after CNS injury, type-1 pericytes differ from scar-forming PDGFRβ + cells.ConclusionsPericyte subpopulations respond differentially to tissue injury, and the production of collagen by type-1 pericytes is organ-dependent. Characterization of the mechanisms underlying scar formation generates cellular targets for future anti-fibrotic therapeutics.Electronic supplementary materialThe online version of this article (doi:10.1186/scrt512) contains supplementary material, which is available to authorized users.

Pulmonary arterial hypertension: yesterday, today, tomorrow.

With more than 25 centuries of efforts to resolve its mysteries, the pulmonary circulation continues to inspire and humble researchers, frustrate clinicians, and scare patients with pulmonary arterial hypertension (PAH). Its mysteries lie in lost manuscripts written in the 12th-century Syria; within the pages of one of the rarest books in history written in 1531 by an author burned alive with his books, condemned by the Inquisition; within a publication from a young urologist self-experimenting 30 years before he received the Nobel prize…What kind of disease is this one that in the age of planning missions to Mars, no medical therapy can save its victims? It is certainly a disease that deserves our respect. The objective of this compendium is to present the state of the art in PAH, but mostly emphasize its future, discussing several evolving theories on its pathogenesis. Its objective is also to comprehensively summarize our current clinical approach to the disease, with patients in mind. In addition to researchers, clinicians, health professionals, students, and residents, its target audience includes patients: the patients who suffer, desperate for answers, and the patients who, one way or another, not only participate in but also fund our research through their taxes and donations. In the age of instant and easy information dissemination, many patients are already quite informed when they visit a specialist. How do we describe the cutting-edge developments in this complex disease to the informed patient? It is not easy, but most would agree that only the expert who really understands his/her subject is able to describe it in lay terms and pass its main messages. Humbled by the complexity of PAH and its challenges, we need to focus on what drives all of us fighting this disease. In addition to our patients, it is scientific curiosity: PAH …

SU‐E‐J‐168: Automated Pancreas Segmentation Based On Dynamic MRI

Purpose:MRI guided radiotherapy is particularly attractive for abdominal targets with low CT contrast. To fully utilize this modality for pancreas tracking, automated segmentation tools are needed. A hybrid gradient, region growth and shape constraint (hGReS) method to segment 2D upper abdominal dynamic MRI is developed for this purpose.Methods:2D coronal dynamic MR images of 2 healthy volunteers were acquired with a frame rate of 5 f/second. The regions of interest (ROIs) included the liver, pancreas and stomach. The first frame was used as the source where the centers of the ROIs were annotated. These center locations were propagated to the next dynamic MRI frame. 4‐neighborhood region transfer growth was performed from these initial seeds for rough segmentation. To improve the results, gradient, edge and shape constraints were applied to the ROIs before final refinement using morphological operations.Results from hGReS and 3 other automated segmentation methods using edge detection, region growth and level set were compared to manual contouring. Results:For the first patient, hGReS resulted in the organ segmentation accuracy as measure by the Dices index (0.77) for the pancreas. The accuracy was slightly superior to the level set method (0.72), and both are significantly more accurate than the edge detection (0.53) and region growth methods (0.42). For the second healthy volunteer, hGReS reliably segmented the pancreatic region, achieving a Dices index of 0.82, 0.92 and 0.93 for the pancreas, stomach and liver, respectively, comparing to manual segmentation. Motion trajectories derived from the hGReS, level set and manual segmentation methods showed high correlation to respiratory motion calculated using a lung blood vessel as the reference while the other two methods showed substantial motion tracking errors. hGReS was 10 times faster than level set.Conclusion:We have shown the feasibility of automated segmentation of the pancreas anatomy based on dynamic MRI.

Influence of intrauterine hypoxia on lung blood vessel development in rats and expression of vascular endothelial growth factor in the lungs

Objective To observe the impact of intrauterine hypoxia on the development of rat lungs and expression of vascular endothelial growth factor (VEGF) in the lungs as the time of hypoxia was extended.Methods To create a model of intrauterine hypoxia,12 pregnant rats were divided into four groups as follows:air-control group,hypoxic 2-day group,hypoxic 6-day group,and hypoxic 10-day group.At birth,we performed pulmonary vascular morphometry in newborn rats with Nis software,and measured pulmonary arterial diameter,wall thickness and wall thickness/pulmonary arterial diameter.We detected expression of VEGF protein by immunohistochemistry and mRNA by real-time polymerase chain reaction.Changes in pulmonary capillary endothelium under electron microscope were observed.One-way analysis of variance and the Student Newman Keuls q (SNK-q) test were applied for statistical analysis.Results As the hypoxic time was extended,wall thickness and wall thickness/pulmonary arterial diameter increased.Compared with the air-control group,pulmonary vascular wall thickness in the hypoxic 10-day group increased [(16.4 ± 5.9) vs (10.8±2.8) μm; q=-8.04,P＜0.05].Wall thickncss/pulmonary artcrial diameter in the hypoxic 10-day group increased compared with that in the air control group,hypoxic 2-day group and hypoxic 6-day group [(31.3±5.1) ％,(22.2±4.9) ％and (23.6±3.9) ％vs (24.1±3.9) ％;q=-7.08,-4.92 and-5.0,all P＜0.05].Expression of VEGF protein in the lungs increased in the hypoxic 6-day group compared with the air-control group [(13.7±3.9) ％ vs (9.3±3.5) ％; q=-6.83,P＜0.05],while the expression was higher in hypoxic 10-day group than in the air-control group and hypoxic 2-day group [(15.2±4.7) ％,(9.3±3.5) ％ vs (11.8 ± 3.3) ％] (q=-9.16 and-5.19,all P＜0.05).Expression of VEGF mRNA in the lungs increased in the hypoxic 6-day group compared with the air-control group [(1.6±0.2)vs (0.8 ±0.2); q=-5.07,P＜0.05],while the expression was higher in the hypoxic 10 day group than in the air-control group and hypoxic 2-day group [(2.2±0.3),(0.8±0.2) vs (1.3±0.2)] (q=-9.54 and-6.42,all P＜0.05).Electron microscopy showed puhnonary capillary endothelial cell swelling as the hypoxic time was extended.In the air-control group,there was no capillary endothelial cell hyperplasia and swelling; in hypoxic 2-day group,there was mild swelling of the capillary endothelial cells and a small amount of hyperplasia; in hypoxic 6-day group,there was moderate swelling of the capillary endothelial cells; and in hypoxic 10-day group:there was significant swelling of the capillary endothelial cells,and pyknosis.Conclusions Intrauterine hypoxia resulted in higher expression of VEGF protcin and mRNA.VEGF in the lungs of newborn rats was involved in the vascular development process. Key words: Fetal hypoxia; Vascular endothelial growth factor A; Lung; Animals, newborn; Rats