cuticleDB: a relational database of Arthropod cuticular proteins

BACKGROUND: The insect exoskeleton or cuticle is a bi-partite composite of proteins and chitin that provides protective, skeletal and structural functions. Little information is available about the molecular structure of this important complex that exhibits a helicoidal architecture. Scores of sequences of cuticular proteins have been obtained from direct protein sequencing, from cDNAs, and from genomic analyses. Most of these cuticular protein sequences contain motifs found only in arthropod proteins. DESCRIPTION: cuticleDB is a relational database containing all structural proteins of Arthropod cuticle identified to date. Many come from direct sequencing of proteins isolated from cuticle and from sequences from cDNAs that share common features with these authentic cuticular proteins. It also includes proteins from the Drosophila melanogaster and the Anopheles gambiae genomes, that have been predicted to be cuticular proteins, based on a Pfam motif (PF00379) responsible for chitin binding in Arthropod cuticle. The total number of the database entries is 445: 370 derive from insects, 60 from Crustacea and 15 from Chelicerata. The database can be accessed from our web server at . CONCLUSIONS: CuticleDB was primarily designed to contain correct and full annotation of cuticular protein data. The database will be of help to future genome annotators. Users will be able to test hypotheses for the existence of known and also of yet unknown motifs in cuticular proteins. An analysis of motifs may contribute to understanding how proteins contribute to the physical properties of cuticle as well as to the precise nature of their interaction with chitin

Structure-Based Discovery of Novel Chemical Classes of Autotaxin Inhibitors

Autotaxin (ATX) is a secreted glycoprotein, widely present in biological fluids, largely responsible for extracellular lysophosphatidic acid (LPA) production. LPA is a bioactive growth-factor-like lysophospholipid that exerts pleiotropic effects in almost all cell types, exerted through at least six G-protein-coupled receptors (LPAR1-6). Increased ATX expression has been detected in different chronic inflammatory diseases, while genetic or pharmacological studies have established ATX as a promising therapeutic target, exemplified by the ongoing phase III clinical trial for idiopathic pulmonary fibrosis. In this report, we employed an in silico drug discovery workflow, aiming at the identification of structurally novel series of ATX inhibitors that would be amenable to further optimization. Towards this end, a virtual screening protocol was applied involving the search into molecular databases for new small molecules potentially binding to ATX. The crystal structure of ATX in complex with a known inhibitor (HA-155) was used as a molecular model docking reference, yielding a priority list of 30 small molecule ATX inhibitors, validated by a well-established enzymatic assay of ATX activity. The two most potent, novel and structurally different compounds were further structurally optimized by deploying further in silico tools, resulting to the overall identification of six new ATX inhibitors that belong to distinct chemical classes than existing inhibitors, expanding the arsenal of chemical scaffolds and allowing further rational design

Interactions between proteins of the endosymbiotic virus of the parasitoid hymenopteran Cotesia congregata and proteins of the lepidopteran immune response

Polydnaviruses (PDVs) are endosymbiotic viruses of parasitoid Hymenoptera. Their particles are injected together with the endoparasitoid’s eggs in the final lepidopteran host, where the viral genes are transcribed. The expressed proteins mediate a series of changes in the host’s physiology, such as developmental delay and immune suppression, thus protecting the hymenopteran embryos from host reactions.The objective of this thesis was to study members of the Ankyrin-repeat protein family, Ank proteins, of the CcBV polydnavirus (Cotesia congregata bracovirus), which lives as a provirus in the hymenopteran species Cotesia congregata, a wasp known to parasitize larvae of the lepidopteran Manduca sexta. CcBV Ank proteins are homologous to ΙκΒ molecules and, therefore, could function as inhibitors of Rel/NF-κΒ transcription factors. Six out of nine CcBV ank genes were subcloned to lepidopteran-specific expression vectors and expressed in insect cells in order to make a comparison among their encoded products. Using transient transfections and subsequent Western immunoblotting, expression levels of the different Ank proteins were found to be unequal, whereas according to immunofluorescence experiments, their subcellular distribution varied among the different family members and often among different cells expressing the same protein. Thus, both the expression levels and the subcellular distribution of CcBV Ank proteins seem to be under regulation.Employing functional assays we tested the effect of Ank proteins on Rel/NF-κΒ transcription factors of the domesticated silkworm Bombyx mori in a lepidopteran cell line. Rel/NF-κΒ factors regulate the transcription of genes implicated in many aspects of the insect immune system, such as antimicrobial (antimicrobial peptide synthesis), antiparasitic (melanization, encapsulation and reactive intermediates production) and antiviral responses through the Imd and Toll signalling pathways. Therefore, it is hypothesized that inhibiting Rel/NF-κΒ factors and the corresponding Imd and Toll pathways of the lepidopteran host’s immune system is crucial for the endoparasitoid’s survival. In order to examine whether CcBV Ank proteins are indeed capable of inhibiting Rel/NF-κΒ factors, and given that the corresponding genes from C. congregata’s natural host M. sexta were unavailable, we used a heterologous system that includes the Trichoplusia ni Hi5 cell line and the transiently expressed silkworm Rel/NF-κΒ transcription factors, Relish1 and RelB. In luciferase assays we examined whether the six viral Ank proteins can suppress the transcription of a firefly luciferase reporter gene placed under the control of a promoter that bears Rel/NF-κΒ binding elements. Five out of the six examined proteins inhibited transcription factor R1d2, a constitutively active Relish1 mutant that participates in the Imd signalling pathway. The inhibition of R1d2 transcriptional activity varied between the different Ank proteins and was dose-dependent for three of them. On the contrary, the transcriptional activity of RelΒ, a factor activated by the Toll pathway, was inhibited only by one out of the six Ank family members. The observed differences between the two signalling pathways suggest that different Ank proteins show specificity against their targets and imply that most CcBV Anks target the Imd pathway, indicating the importance of this pathway in anti-viral responses. Immunofluorescence experiments in Hi5 cells transiently over-expressing the viral proteins and the silkworm transcription factors revealed nuclear co-localization between transcription factor R1d2 and Ank proteins, while RelΒ co-localized with its Ank2 inhibitor in the cytoplasm. Moreover, two Ank proteins were found to interact with full-length Relish1 in pull-down experiments which, together with the R1d2 inhibition observed in the functional assays, implies that Ank proteins could act as Relish1 inhibitors in vivo.Furthermore, the possible suppression of mammalian Rel/NF-κB factors by CcBV Ank proteins was also tested. Six ank genes were subcloned in an expression vector suitable for expression in mammalian cells and the effect of the corresponding proteins on endogenous transcription factor NF-κB of HEK293 cells was examined. Tumor necrosis factor α (TNF-α) was used as an inducer of the endogenous mammalian signalling pathway that is initiated by the TNF-receptor and that corresponds to the insect Imd pathway. Our results showed that two members of the Ank protein family inhibit the TNFR pathway in HEK293 cells. Conclusively, this thesis describes the first attempt to compare multiple members of a PDV Ank family both at expression/localization level and functional level, thus giving the opportunity to comprehend the existing variation in PDV protein families. The results are in agreement with the hypothesis that polydnavirus Ankyrin-repeat proteins act as Rel/NF-κΒ inhibitors; both in lepidopteran insect cells and in mammalian cells. In fact, Ank proteins are capable of suppressing both major immune signalling pathways, Toll and Imd, in insect cells. Finally, it is evident that Ank proteins of a PDV endosymbiotic of a certain hymenopteran species can block immune regulators of the natural lepidopteran host, as well as of other Lepidoptera. Therefore, PDV proteins, like Anks, could be employed for the immune suppression of a range of insects in order to enhance the effectiveness of pest control applications involving entomopathogenic viruses, bacteria or fungi.Οι polydna ιοί είναι μια ομάδα ιών που ενδοσυμβιώνει με ενδοπαρασιτοειδή υμενόπτερα έντομα (σφήκες) και συμμετέχει στον παρασιτισμό λεπιδοπτέρων εντόμων από τα Υμενόπτερα. Τα σωματίδια των ιών ενίονται μαζί με τα γονιμοποιημένα αυγά των σφηκών στα λεπιδόπτερα έντομα-ξενιστές. Τα γονίδια των polydna ιών εκφράζονται στα κύτταρα του λεπιδόπτερου ξενιστή και οι παραγόμενες πρωτεΐνες επιδρούν στη φυσιολογία του ξενιστή καθυστερώντας την ανάπτυξή του και καταστέλλοντας το ανοσοποιητικό του σύστημα. Με αυτόν τον τρόπο τα αυγά και τα έμβρυα της σφήκας προστατεύονται από αντιδράσεις άμυνας του ξενιστή και μπορούν να αναπτυχθούν. Στόχος της διατριβής ήταν η μελέτη μελών της οικογένειας πρωτεϊνών που περιέχουν επαναλήψεις αγκυρίνης (πρωτεΐνες Ank) του polydna ιού CcBV (Cotesia congregata bracovirus), ο οποίος ενδοσυμβιώνει με τη σφήκα Cotesia congregata και συμμετέχει στον παρασιτισμό των προνυμφών του Λεπιδοπτέρου Manduca sexta. Οι πρωτεΐνες Ank του ιού CcBV είναι ομόλογες με μόρια ΙκΒ, δηλαδή δυνητικοί αναστολείς μεταγραφικών παραγόντων τύπου Rel/NF-κΒ. Έξι γονίδια ank του CcBV υποκλωνοποιήθηκαν σε φορείς κατάλληλους για έκφραση σε κύτταρα λεπιδοπτέρων εντόμων ώστε να γίνει συγκριτική μελέτη των ιδιοτήτων των κωδικοποιούμενων πρωτεϊνών. Με παροδικές διαμολύνσεις και ανοσοαποτυπώσεις κατά Western βρέθηκε ότι τα διάφορα μέλη της οικογένειας παρουσιάζουν άνισα επίπεδα έκφρασης, ενώ πειράματα ανοσοφθορισμού ανέδειξαν ετερογένεια στον υποκυτταρικό εντοπισμό τους, με κάποιες πρωτεΐνες να απαντώνται σε περισσότερα του ενός υποκυτταρικά διαμερίσματα σε διαφορετικά κύτταρα, γεγονός που υποδηλώνει ότι ο εντοπισμός και η έκφρασή τους υπόκεινται σε ρύθμιση.Με χρήση κατάλληλων λειτουργικών πειραμάτων, εξετάστηκε επίσης η επίδραση των πρωτεϊνών Ank στην ενεργότητα Rel/NF-κΒ παραγόντων του καλλιεργούμενου μεταξοσκώληκα Bombyx mori σε κυτταρικές σειρές λεπιδοπτέρων εντόμων. Οι παράγοντες Rel/NF-κΒ ρυθμίζουν, μέσω των οδών μεταγωγής σήματος Imd και Toll, την έκφραση γονιδίων που ελέγχουν πολλαπλές πτυχές του ανοσοποιητικού συστήματος των εντόμων, όπως την παραγωγή αντιμικροβιακών πεπτιδίων, καθώς και αποκρίσεις που αφορούν πιο άμεσα την αντιμετώπιση του παρασιτισμού, όπως η μελανοποίηση, η εγκύστωση, η παραγωγή ελευθέρων ριζών και οι αντι-ιικές αποκρίσεις. Επομένως, πιθανολογείται ότι η αναστολή των παραγόντων Rel/NF-κΒ και η συνεπαγόμενη παρεμπόδιση των σηματοδοτικών οδών Toll και Imd του ανοσοποιητικού συστήματος των λεπιδοπτέρων ξενιστών από πρωτεΐνες των polydna ιών, και ειδικότερα από τις πρωτεΐνες Ank, είναι σημαντική για την επιβίωση των παρασιτοειδών. Για να διαπιστωθεί αν η συγκεκριμένη υπόθεση ευσταθεί και με δεδομένη τη μη διαθεσιμότητα των γονιδίων Rel/NF-κΒ παραγόντων του φυσικού ξενιστή της C. congregata, M. sexta, χρησιμοποιήθηκε ένα ετερόλογο λειτουργικό σύστημα που περιέλαβε τα γονίδια των ετερόλογων μεταγραφικών παραγόντων Relish1 και RelB του B. mori, που συμμετέχουν στις οδούς μεταγωγής σήματος Imd και Toll, αντίστοιχα, σε συνδυασμό με την κυτταρική σειρά Hi5 του Λεπιδοπτέρου Trichoplusia ni. Σε λειτουργικές δοκιμασίες φωταύγειας διερευνήθηκε η επίδραση των έξι ιικών πρωτεϊνών Ank στη μεταγραφή ενός γονιδίου αναφοράς firefly luciferase, που είχε τεθεί υπό τον έλεγχο υποκινητών με στοιχεία πρόσδεσης Rel/NF-κΒ για τους δύο μεταγραφικούς παράγοντες του μεταξοσκώληκα. Τα αποτελέσματα έδειξαν ότι οι πέντε από τις έξι πρωτεΐνες του CcBV που ελέγχθηκαν προκάλεσαν αναστολή στην ενεργότητα του μεταγραφικού παράγοντα R1d2, μια συστατικά ενεργή μορφή του Relish1, στο μονοπάτι Imd, με την αναστολή να είναι διαφορική για τα έξι μέλη και δοσοεξαρτόμενη για τρία μέλη της οικογένειας που εξετάστηκαν σε σχετικά πειράματα δοσοεξάρτησης. Αντίθετα, η ενεργότητα του μεταγραφικού παράγοντα RelΒ, που ενεργοποιείται από το μονοπάτι Toll, βρέθηκε να αναστέλλεται μόνο από ένα από τα έξι μέλη της οικογένειας Ank που εξετάστηκαν. Η διαφοροποίηση ανάμεσα στις δύο οδούς μεταγωγής σήματος υποδηλώνει την εξειδίκευση των διαφορετικών ιικών πρωτεϊνών Ank ως προς τα μόρια-στόχους τους και υποδεικνύει ότι τα περισσότερα μέλη στοχεύουν την οδό μεταγωγής σήματος Imd, γεγονός πιθανά ενδεικτικό του ρόλου που η συγκεκριμένη οδός διαδραματίζει στην αντι-ιική προστασία των εντόμων. Σε κύτταρα Hi5 που υπερεκφράζουν παροδικά τις ιικές πρωτεΐνες και τους μεταγραφικούς παράγοντες παρατηρήθηκε πυρηνικός συνεντοπισμός του μεταγραφικού παράγοντα R1d2 με αρκετές από τις πρωτεΐνες Ank που αναστέλλουν τη λειτουργία του, ενώ ο παράγοντας RelΒ παρουσίασε συνεντοπισμό με την πρωτεΐνη-αναστολέα του στο κυτταρόπλασμα. Επιπλέον, πειράματα συγκατακρήμνισης έδειξαν αλληλεπίδραση ανάμεσα σε δύο πρωτεΐνες Ank και τον πλήρους μήκους παράγοντα Relish1, γεγονός που, σε συνδυασμό με την αναστολή του R1d2 από τις ιικές πρωτεΐνες, υποδηλώνει ότι οι πρωτεΐνες Ank πιθανώς δρουν ως αναστολείς του Relish1 και in vivo.Στα πλαίσια της προσπάθειας για διαλεύκανση πιθανής δράσης των πρωτεϊνών Ank του CcBV σε Rel/NF-κB παράγοντες θηλαστικών, τα έξι ank γονίδια υποκλωνοποιήθηκαν σε φορέα κατάλληλο για έκφραση σε κύτταρα θηλαστικών και εξετάστηκε η δράση των αντίστοιχων πρωτεϊνών στη λειτουργικότητα του ενδογενούς μεταγραφικού παράγοντα NF-κB κυττάρων HEK293. Χρησιμοποιώντας την κυτταροκίνη TNF-α ως επαγωγέα του NF-κB, διαπιστώθηκε η ανασταλτική δράση δύο πρωτεϊνών στην ανοσολογική οδό μεταγωγής σήματος των θηλαστικών που ενεργοποιείται μέσω του υποδοχέα του παράγοντα νέκρωσης όγκων TNFR και είναι αντίστοιχη με την Imd οδό των εντόμων. Συμπεραματικά, με τις μελέτες που περιγράφονται σε αυτή τη διατριβή, επιχειρήθηκε για πρώτη φορά η σύγκριση ανάμεσα σε πολλά μέλη μιας οικογένειας πρωτεϊνών Ank των ενδοσυμβιωτικών polydna ιών σε επίπεδο έκφρασης και υποκυτταρικής κατανομής αλλά και σε λειτουργικό επίπεδο δίνοντας έτσι τη δυνατότητα κατανόησης της λειτουργικής ποικιλομορφίας που υπάρχει σε πρωτεϊνικές οικογένειες των polydna ιών. Τα αποτελέσματα είναι συμβατά με την υπόθεση ότι οι πρωτεΐνες επαναλήψεων αγκυρίνης των polydna ιών δρουν ως αναστολείς παραγόντων Rel/NF-κΒ, τόσο στα λεπιδόπτερα έντομα όσο και στα θηλαστικά. Μάλιστα, στα έντομα, οι πρωτεΐνες Ank έχουν τη δυνατότητα να καταστέλλουν και τις δύο κύριες ανοσολογικές οδούς μεταγωγής σήματος, Toll και Imd. Επίσης, φαίνεται ότι τα μέλη της οικογένειας Ank του ενδοσυμβιωτικού ιού ενός Υμενοπτέρου που παρασιτεί ένα συγκεκριμένο Λεπιδόπτερο μπορούν να παρεμποδίζουν τη λειτουργία ρυθμιστών της ανοσίας και άλλων Λεπιδοπτέρων πέραν του φυσικού του ξενιστή. Επομένως, πρωτεΐνες των polydna ιών, όπως οι Ank, θα μπορούσαν να χρησιμοποιηθούν για την καταστολή του ανοσοποιητικού συστήματος των εντόμων στα πλαίσια ενίσχυσης ήδη υπάρχοντων στρατηγικών καταπολέμησης επιβλαβών εντόμων, οι οποίες βασίζονται στη χρήση εντομοπαθογόνων μικροοργανισμών

Autotaxin in Pathophysiology and Pulmonary Fibrosis

Lysophospholipid signaling is emerging as a druggable regulator of pathophysiological responses, and especially fibrosis, exemplified by the relative ongoing clinical trials in idiopathic pulmonary fibrosis (IPF) patients. In this review, we focus on ectonucleotide pyrophosphatase-phosphodiesterase 2 (ENPP2), or as more widely known Autotaxin (ATX), a secreted lysophospholipase D (lysoPLD) largely responsible for extracellular lysophosphatidic acid (LPA) production. In turn, LPA is a bioactive phospholipid autacoid, forming locally upon increased ATX levels and acting also locally through its receptors, likely guided by ATX's structural conformation and cell surface associations. Increased ATX activity levels have been detected in many inflammatory and fibroproliferative conditions, while genetic and pharmacologic studies have confirmed a pleiotropic participation of ATX/LPA in different processes and disorders. In pulmonary fibrosis, ATX levels rise in the broncheoalveolar fluid (BALF) and stimulate LPA production. LPA engagement of its receptors activate multiple G-protein mediated signal transduction pathways leading to different responses from pulmonary cells including the production of pro-inflammatory signals from stressed epithelial cells, the modulation of endothelial physiology, the activation of TGF signaling and the stimulation of fibroblast accumulation. Genetic or pharmacologic targeting of the ATX/LPA axis attenuated disease development in animal models, thus providing the proof of principle for therapeutic interventions

Lysophosphatidic Acid Is a Proinflammatory Stimulus of Renal Tubular Epithelial Cells

Chronic kidney disease (CKD) refers to a spectrum of diseases defined by renal fibrosis, permanent alterations in kidney structure, and low glomerular-filtration rate. Prolonged epithelial-tubular damage involves a series of changes that eventually lead to CKD, highlighting the importance of tubular epithelial cells in this process. Lysophosphatidic acid (LPA) is a bioactive lipid that signals mainly through its six cognate LPA receptors and is implicated in several chronic inflammatory pathological conditions. In this report, we have stimulated human proximal tubular epithelial cells (HKC-8) with LPA and 175 other possibly pathological stimuli, and simultaneously detected the levels of 27 intracellular phosphoproteins and 32 extracellular secreted molecules with multiplex ELISA. This quantification revealed a large amount of information concerning the signaling and the physiology of HKC-8 cells that can be extrapolated to other proximal tubular epithelial cells. LPA responses clustered with pro-inflammatory stimuli such as TNF and IL-1, promoting the phosphorylation of important inflammatory signaling hubs, including CREB1, ERK1, JUN, IκΒα, and MEK1, as well as the secretion of inflammatory factors of clinical relevance, including CCL2, CCL3, CXCL10, ICAM1, IL-6, and IL-8, most of them shown for the first time in proximal tubular epithelial cells. The identified LPA-induced signal-transduction pathways, which were pharmacologically validated, and the secretion of the inflammatory factors offer novel insights into the possible role of LPA in CKD pathogenesis

Autotaxin Activity in Chronic Subdural Hematoma: A Prospective Clinical Study

Autotaxin (ATX) is the ectoenzyme producing the bulk of lysophosphatidic acid (LPA) in circulation. ATX and LPA-mediated signaling (the ATX-LPA axis) play critical roles in the vascular and nervous system development. In adults, this axis contributes to diverse processes, including coagulation, inflammation, fibroproliferation and angiogenesis under physiological and/or pathophysiological conditions. Given evidence implicating several of these processes in chronic subdural hematoma (CSDH) pathogenesis and development, we assessed ATX activity in CSDH patients. Twenty-eight patients were recruited. Blood and hematoma fluid were collected. Enzymatic assays were used to establish serum and hematoma ATX activity. Enzyme-linked immunosorbent assays were used to establish hematoma beta trace (BT) levels, a cerebrospinal fluid (CSF) marker, in a hematoma. ATX activity was nearly three folds higher in hematoma compared to serum (P < 0.001). There was no significant correlation between BT levels and ATX activity in a hematoma. The present results show, for the first time, that ATX is catalytically active in the hematoma fluid of CSDH patients. Moreover, our findings of significantly elevated ATX activity in hematoma compared to serum, implicate the ATX-LPA axis in CSDH pathophysiology. The CSF origin of ATX could not be inferred with the present results. Additional research is warranted to establish the significance of the ATX-LPA axis in CSDH and its potential as a biomarker and/or therapeutic target

Autotaxin and Endotoxin-Induced Acute Lung Injury.

Acute Lung Injury (ALI) is a life-threatening, diffuse heterogeneous lung injury characterized by acute onset, pulmonary edema and respiratory failure. Lipopolysaccharide (LPS) is a common cause of both direct and indirect lung injury and when administered to a mouse induces a lung phenotype exhibiting some of the clinical characteristics of human ALI. Here, we report that LPS inhalation in mice results in increased bronchoalveolar lavage fluid (BALF) levels of Autotaxin (ATX, Enpp2), a lysophospholipase D largely responsible for the conversion of lysophosphatidylcholine (LPC) to lysophosphatidic acid (LPA) in biological fluids and chronically inflamed sites. In agreement, gradual increases were also detected in BALF LPA levels, following inflammation and pulmonary edema. However, genetic or pharmacologic targeting of ATX had minor effects in ALI severity, suggesting no major involvement of the ATX/LPA axis in acute inflammation. Moreover, systemic, chronic exposure to increased ATX/LPA levels was shown to predispose to and/or to promote acute inflammation and ALI unlike chronic inflammatory pathophysiological situations, further suggesting a differential involvement of the ATX/LPA axis in acute versus chronic pulmonary inflammation

Genetic deletion of Autotaxin from CD11b+ cells decreases the severity of experimental autoimmune encephalomyelitis.

Autotaxin (ATX) is a secreted lysophospholipase D catalyzing the extracellular production of lysophosphatidic acid (LPA), a growth factor-like signaling lysophospholipid. ATX and LPA signaling have been incriminated in the pathogenesis of different chronic inflammatory diseases and various types of cancer. In this report, deregulated ATX and LPA levels were detected in the spinal cord and plasma of mice during the development of experimental autoimmune encephalomyelitis (EAE). Among the different sources of ATX expression in the inflamed spinal cord, F4/80+ CD11b+ cells, mostly activated macrophages and microglia, were found to express ATX, further suggesting an autocrine role for ATX/LPA in their activation, an EAE hallmark. Accordingly, ATX genetic deletion from CD11b+ cells attenuated the severity of EAE, thus proposing a pathogenic role for the ATX/LPA axis in neuroinflammatory disorders

Deletion of ATX from myeloid cells had no effects in ALI development.

<p>Aerosolized LPS was administered in mice where ATX was deleted from the myeloid cells (LysMEnpp2^-/-) and wild type littermate mice. Mice were sacrificed 24 hours later. A-C. Histological analysis (A) and BALF measurements (B & C) showed no differences in inflammation and edema between mice lacking ATX expression from myeloid cells and their wild type littermates. D. Conditional deletion of ATX from myeloid cells had no effects in BALF ATX activity.</p

Pharmacologic inhibition of ATX has no effects in ALI development.

<p>GWJ-A-23 was injected intraperitoneally before challenging mice with aerosolized LPS. Mice were sacrificed 24 hours later (n = 3–7; a representative experiment out of two is shown). A-C. Histological analysis (A) and indicated BALF measurements (B,C) suggested minor effects in ALI development, despite decreased BALF ATX activity (D) and the corresponding BALF LPA levels (E).</p