Docosatetraenoyl LPA is elevated in exhaled breath condensate in idiopathic pulmonary fibrosis

Background: Idiopathic pulmonary fibrosis (IPF) is a progressive and fatal disease with no effective medical therapies. Recent research has focused on identifying the biological processes essential to the development and progression of fibrosis, and on the mediators driving these processes. Lysophosphatidic acid (LPA), a biologically active lysophospholipid, is one such mediator. LPA has been found to be elevated in bronchoalveolar lavage (BAL) fluid of IPF patients, and through interaction with its cell surface receptors, it has been shown to drive multiple biological processes implicated in the development of IPF. Accordingly, the first clinical trial of an LPA receptor antagonist in IPF has recently been initiated. In addition to being a therapeutic target, LPA also has potential to be a biomarker for IPF. There is increasing interest in exhaled breath condensate (EBC) analysis as a non-invasive method for biomarker detection in lung diseases, but to what extent LPA is present in EBC is not known. Methods: In this study, we used liquid chromatography-tandem mass spectrometry (LC-MS/MS) to assess for the presence of LPA in the EBC and plasma from 11 IPF subjects and 11 controls. Results: A total of 9 different LPA species were detectable in EBC. Of these, docosatetraenoyl (22:4) LPA was significantly elevated in the EBC of IPF subjects when compared to controls (9.18 pM vs. 0.34 pM; p = 0.001). A total of 13 different LPA species were detectable in the plasma, but in contrast to the EBC, there were no statistically significant differences in plasma LPA species between IPF subjects and controls. Conclusions: These results demonstrate that multiple LPA species are detectable in EBC, and that 22:4 LPA levels are elevated in the EBC of IPF patients. Further research is needed to determine the significance of this elevation of 22:4 LPA in IPF EBC, as well as its potential to serve as a biomarker for disease severity and/or progression

Сорбционно-структурные свойства аэрогельных материалов на основе биополимеров

В настоящее время для выведения избыточного количества тяжелых металлов и токсинов из живых организмов успешно применяются аэрогельные материалы в качестве энтеро- и апликационных сорбентов. Неисчерпаемой сырьевой базой для создания аэрогельных материалов являются природные биополимеры альгинат и хитозан, а также различные производные лигнина. На их основе разработано значительное количество сорбционных материалов и раневых покрытий различных типов, что связано не только с широким спектром физико-химических свойств названных полимеров и их уже доказанной медико-биологической активностью, но и с распространенностью и возобновляемостью сырьевых источников для производства данных полимеров, простотой извлечения, возможностью достижения высокой степени очистки и сравнительно невысокой ценой. Ключевой стадией синтеза аэрогельных материалов является формирование прочного гидрогеля – каркаса. Один из технологических приемов – получение интерполиэлектролитного армирующего гидрогеля. В работе предложены 2 упаковочные модели формирования структуры интерполиэлектролитных комплексов на основе пар биополимеров: «альгинат натрия – хитозан» и «лигносульфонат натрия – хитозан». Первая модель – блочная, при которой структура формируется за счет ионных связей между карбоксильными группами альгината натрия и аминогруппами хитозана, а также кооперативной системы водородных связей и дисперсионных взаимодействий. Вторая модель – агрегационно-трубчатая, структура которой образуется посредством ионных связей между сульфогруппами (в составе палочкообразных надмолекулярных структур лигносульфоната натрия) и аминогруппами хитозана, а также водородных связей и дисперсионных взаимодействий. При высушивании интерполиэлектролитных комплексов в сверхкритических условиях формируются прочные фазовые контакты, при этом изменения в структуре геля становятся необратимыми. В результате получены гидрофобные микро- и мезопористые 2-компонентные аэрогельные материалы, различающиеся внутренней структурой. Аэрогельные материалы, структура которых образована по 1-й из названных моделей, характеризуются фибриллярной структурой, а по 2-й ‒ структурными элементами сферической формы. Полученные аэрогельные материалы обладают высокой сорбционной активностью по отношению к воде и широкому кругу тяжелых металлов и низкомолекулярных токсинов. Цель работы – исследование структурно-сорбционных свойств аэрогельных материалов, основа которых – биополимеры различной структурной организации. Значительное увеличение сорбционной активности аэрогельных материалов «альгинат натрия – хитозан» в сравнении с «лигносульфонат натрия – хитозан» связано, по-видимому, с их различной надмолекулярной структурой. Действует совокупность механизмов сорбции: намокание, всасывание, диффузия, осмотические явления и химическое взаимодействие, обусловленное высокопористой структурой аэрогельных материалов и наличием сорбционно-активных центров. Для цитирования: Бровко О.С., Паламарчук И.А., Горшкова Н.А., Богданович Н.И., Ивахнов А.Д. Сорбционно-структурные свойства аэрогельных материалов на основе биополимеров // Изв. вузов. Лесн. журн. 2023. № 6. С. 190–203. https://doi.org/10.37482/0536-1036-2023-6-190-20

Role of Nox4 and Nox2 in Hyperoxia-Induced Reactive Oxygen Species Generation and Migration of Human Lung Endothelial Cells

Abstract In vascular endothelium, the major research focus has been on reactive oxygen species (ROS) derived from Nox2. The role of Nox4 in endothelial signal transduction, ROS production, and cytoskeletal reorganization is not well defined. In this study, we show that human pulmonary artery endothelial cells (HPAECs) and human lung microvascular endothelial cells (HLMVECs) express higher levels of Nox4 and p22phox compared to Nox1, Nox2, Nox3, or Nox5. Immunofluorescence microscopy and Western blot analysis revealed that Nox4 and p22phox, but not Nox2 or p47phox, are localized in nuclei of HPAECs. Further, knockdown of Nox4 with siRNA decreased Nox4 nuclear expression significantly. Exposure of HPAECs to hyperoxia (3-24h) enhanced mRNA and protein expression of Nox4, and Nox4 siRNA decreased hyperoxia-induced ROS production. Interestingly, Nox4 or Nox2 knockdown with siRNA upregulated the mRNA and protein expression of the other, suggesting activation of compensatory mechanisms. A similar upregulation of Nox4 mRNA was observed in Nox2 2/ko mice. Downregulation of Nox4, or pretreatment with N-acetylcysteine, attenuated hyperoxia-induced cell migration and capillary tube formation, suggesting that ROS generated by Nox4 regulate endothelial cell motility. These results indicate that Nox4 and Nox2 play a physiological role in hyperoxia-induced ROS production and migration of ECs. Antioxid. Redox Signal. 11, 747-764.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/78121/1/ars.2008.2203.pd

Intracellular S1P Generation Is Essential for S1P-Induced Motility of Human Lung Endothelial Cells: Role of Sphingosine Kinase 1 and S1P Lyase

Earlier we have shown that extracellular sphingosine-1-phosphate (S1P) induces migration of human pulmonary artery endothelial cells (HPAECs) through the activation of S1P(1) receptor, PKCε, and PLD2-PKCζ-Rac1 signaling cascade. As endothelial cells generate intracellular S1P, here we have investigated the role of sphingosine kinases (SphKs) and S1P lyase (S1PL), that regulate intracellular S1P accumulation, in HPAEC motility

Lysophosphatidic acid-induced transactivation of epidermal growth factor receptor regulates cyclo-oxygenase-2 expression and prostaglandin E2release via C/EBPβ in human bronchial epithelial cells

Detail, section through pedimented lid, Alexander Sarcophagus (plate #30); Fragments d'architecture antique d'après les relevés & restaurations des anciens pensionnaires de l'Académie de France à Rome; publiés sous la direction de H. d'Espony ...Publication info: Paris, C. Schmid [189-?]-1905.Physical descrip: 2 v. 200 pl. (incl. plans, diagrs.) Source: University of Toronto Libraries; http://main.library.utoronto.ca/ (accessed 1/12/2008

Properties of Fluorescent Far-Red Anti-TNF Nanobodies

Upregulation of the expression of tumor necrosis factor (TNF-α, TNF) has a significant role in the development of autoimmune diseases. The fluorescent antibodies binding TNF may be used for personalized therapy of TNF-dependent diseases as a tool to predict the response to anti-TNF treatment. We generated recombinant fluorescent proteins consisting of the anti-TNF module based on the variable heavy chain (VHH) of camelid antibodies fused with the far-red fluorescent protein Katushka (Kat). Two types of anti-TNF VHH were developed: one (BTN-Kat) that was bound both human or mouse TNF, but did not neutralize their activity, and a second (ITN-Kat) that was binding and neutralizing human TNF. BTN-Kat does not interfere with TNF biological functions and can be used for whole-body imaging. ITN-Kat can be evaluated in humanized mice or in cells isolated from humanized mice. It is able to block human TNF (hTNF) activities both in vitro and in vivo and may be considered as a prototype of a theranostic agent for autoimmune diseases

Photolysis of caged sphingosine-1-phosphate induces barrier enhancement and intracellular activation of lung endothelial cell signaling pathways

Sphingosine-1-phosphate (S1P) is a bioactive sphingolipid that mediates cellular functions by ligation via G protein-coupled S1P receptors. In addition to its extracellular action, S1P also has intracellular effects; however, the signaling pathways modulated by intracellular S1P remain poorly defined. We have previously demonstrated a novel pathway of intracellular S1P generation in human lung endothelial cells (ECs). In the present study, we examined the role of intracellular S1P generated by photolysis of caged S1P on EC barrier regulation and signal transduction. Intracellular S1P released from caged S1P caused mobilization of intracellular calcium, induced activation of MAPKs, redistributed cortactin, vascular endothelial cadherin, and β-catenin to cell periphery, and tightened endothelial barrier in human pulmonary artery ECs. Treatment of cells with pertussis toxin (PTx) had no effect on caged S1P-mediated effects on Ca2+ mobilization, reorganization of cytoskeleton, cell adherens junction proteins, and barrier enhancement; however, extracellular S1P effects were significantly attenuated by PTx. Additionally, intracellular S1P also activated small GTPase Rac1 and its effector Ras GTPase-activating-like protein IQGAP1, suggesting involvement of these proteins in the S1P-mediated changes in cell-to-cell adhesion contacts. Downregulation of sphingosine kinase 1 (SphK1), but not SphK2, with siRNA or inhibition of SphK activity with an inhibitor 2-(p-hydroxyanilino)-4-(p-chlorophenyl) thiazole (CII) attenuated exogenously administrated S1P-induced EC permeability. Furthermore, S1P1 receptor inhibitor SB649164 abolished exogenous S1P-induced transendothelial resistance changes but had no effect on intracellular S1P generated by photolysis of caged S1P. These results provide evidence that intracellular S1P modulates signal transduction in lung ECs via signaling pathway(s) independent of S1P receptors