Solution structure of the Legionella pneumophila Mip-rapamycin complex

<p>Abstract</p> <p>Background</p> <p><it>Legionella pneumphila </it>is the causative agent of Legionnaires' disease. A major virulence factor of the pathogen is the homodimeric surface protein Mip. It shows peptidyl-prolyl cis/trans isomerase activty and is a receptor of FK506 and rapamycin, which both inhibit its enzymatic function. Insight into the binding process may be used for the design of novel Mip inhibitors as potential drugs against Legionnaires' disease.</p> <p>Results</p> <p>We have solved the solution structure of free Mip^77–213and the Mip^77–213-rapamycin complex by NMR spectroscopy. Mip^77–213showed the typical FKBP-fold and only minor rearrangements upon binding of rapamycin. Apart from the configuration of a flexible hairpin loop, which is partly stabilized upon binding, the solution structure confirms the crystal structure. Comparisons to the structures of free FKBP12 and the FKBP12-rapamycin complex suggested an identical binding mode for both proteins.</p> <p>Conclusion</p> <p>The structural similarity of the Mip-rapamycin and FKBP12-rapamycin complexes suggests that FKBP12 ligands may be promising starting points for the design of novel Mip inhibitors. The search for a novel drug against Legionnaires' disease may therefore benefit from the large variety of known FKBP12 inhibitors.</p

Mechanistic interrogation of combination Bevacizumab/dual PI3K/mTOR inhibitor response in Glioblastoma implementing novel MR and PET imaging biomarkers.

Purpose: Resistance to bevacizumab (BEV) in glioblastoma (GBM) is believed to occur via activation of molecular networks including the mTOR/PI3K pathway. Implementing an MRI/PET molecular imaging biomarker approach, we sought to interrogate response to combining BEV with the mTOR/PI3K inhibitor BEZ235. Methods: Tumors were established by orthotopically implanting U87MG-luc2 in mice. Animals were treated with BEZ235 and/or BEV, and imaged using diffusion weighted-MRI, T2 weighted (T2w), and T2* weighted (T2*w) before and following delivery of superparamagnetic iron oxide (SPIO) contrast. Maps for changes in relaxation rates: ΔR2, ΔR2* and apparent diffusion coefficient (ADC) were calculated. Vessel Size Index (VSI) and micro vessel density index (MDI) were derived. 3´-deoxy-3´-[18F]fluorothymidine ([18F]FLT)- and O-(2-[18F]fluoroethyl)-L-tyrosine ([18F]FET) PET was further performed and tumor endothelium/proliferation markers assessed by immunohistochemistry. Results: Treatment with BEV resulted in a pronounced decrease in tumor volume (T2w MRI). No additive effect on tumour volume was observed in BEV/BEZ235 combination compared with BEV monotherapy. Ki67 proliferation index staining and [18F]FLT uptake studies were used to support observations. Using ΔR2* and ΔR2 values respectively, BEZ235 + BEV combination significantly reduced tumor microvessel volume in comparison to BEV alone. Decreased MDI was further observed in the combination group; supported by von Willebrand Factor (vWF) immunohistochemistry. We observed decreased [18F]FET uptake following BEV, but failed to observe further reduced [18F]FET uptake in the combination cohort. vWF IHC analysis showed mean tumor vessel size increased in all cohorts. Conclusions: Assessing MR imaging biomarker parameters together with [18F]FET and [18F]FLT PET, informed drug combination mechanism of action and provided clues as to potential clinical response. Translation of a BEZ35/BEV combination regimen could support reduction of peritumoral edemaobviating the requirement for steroids. Implementing hypothesis driven molecular imaging studies facilitates the interrogation of drug response in the pre-clinic. These data may more accurately predict the clinical potential of novel therapeutic approaches in oncology

ARTEFACTS: How do we want to deal with the future of our one and only planet?

The European Commission’s Science and Knowledge Service, the Joint Research Centre (JRC), decided to try working hand-in-hand with leading European science centres and museums. Behind this decision was the idea that the JRC could better support EU Institutions in engaging with the European public. The fact that European Union policies are firmly based on scientific evidence is a strong message which the JRC is uniquely able to illustrate. Such a collaboration would not only provide a platform to explain the benefits of EU policies to our daily lives but also provide an opportunity for European citizens to engage by taking a more active part in the EU policy making process for the future. A PILOT PROGRAMME To test the idea, the JRC launched an experimental programme to work with science museums: a perfect partner for three compelling reasons. Firstly, they attract a large and growing number of visitors. Leading science museums in Europe have typically 500 000 visitors per year. Furthermore, they are based in large European cities and attract local visitors as well as tourists from across Europe and beyond. The second reason for working with museums is that they have mastered the art of how to communicate key elements of sophisticated arguments across to the public and making complex topics of public interest readily accessible. That is a high-value added skill and a crucial part of the valorisation of public-funded research, never to be underestimated. Finally museums are, at present, undergoing something of a renaissance. Museums today are vibrant environments offering new techniques and technologies to both inform and entertain, and attract visitors of all demographics.JRC.H.2-Knowledge Management Methodologies, Communities and Disseminatio

Palliative Chemotherapy of the non-small-cell lung cancer

In der vorliegenden Studie wurde die Effektivität palliativer Therapieansätze für Patienten mit fortgeschrittenem nicht-kleinzelligem Bronchialkarzinom untersucht. Zugrunde liegen die Daten von 75 Patienten, die an der Medizinischen Universitätsklinik Würzburg im Schwerpunkt Pneumologie zwischen Januar 1995 und Juni 1997 erstmalig eine Chemo- bzw. Radiochemotherapie bei inoperablem Bronchialkarzinom erhielten. Die allgemeinen Patientenmerkmale des untersuchten Probandenkollektivs, wie Alter, Geschlecht, Rauchverhalten, Histologie, Lokalisation und Stadium des Tumors (nach der Stadieneinteilung durch die TNM-Klassifikation), sowie der Allgemeinzustand (nach Karnofsky) sind durchaus repräsentativ verteilt und legen somit die Basis zur Beurteilung der Ergebnisse dieser Studie. Mittels nichtparametrischer Gruppentests wurden zum einen der Therapieerfolg und das Überleben taxanhaltiger Therapien mit denen taxanfreier Behandlungen verglichen, zum anderen wurde im gleichen Patientenkollektiv die Effektivität der Radiochemotherapien mit der von Therapien ohne Radiatio analysiert. Ein Kriterium zur Bewertung der unterschiedlichen Behandlungsansätze ist die Therapiebelastung für den Patienten. Hier zeigt sich trotz häufiger auftretenden Nebenwirkungen der Therapien mit Taxan, eine deutlich geringere Abbruchrate als bei taxanfreien Therapien, und damit eine bessere Durchführbarkeit dieses Ansatzes. Vergleicht man Radiochemotherapien mit Behandlungen ohne Strahlentherapie, so schneiden Radiochemotherapien in Bezug auf die Abbruchrate deutlich besser ab, obwohl Nebenwirkungen bei diesen häufiger sind. Allerdings verweigern Patienten mit kombinierter Radiochemotherapie öfter die Behandlung, scheinbar sind die unerwünschten Wirkungen dieser Behandlung subjektiv belastender. Insgesamt ist das Nebenwirkungsprofil der angewendeten Therapieansätze mit den Literaturangaben durchaus vergleichbar, wenn auch im eigenen Patientengut bei taxanhaltigen Behandlungen deutlich weniger nicht hämatologische Nebenwirkungen und insgesamt häufiger Anämien auftreten. Die mediane Überlebenszeit aller Patienten lag bei 10 Monaten; die Überlebensrate nach einem Jahr beträgt 40,5 %. Im Vergleich mit Angaben aus der Literatur sind diese Ergebnisse durchaus repräsentativ. Sowohl die Ansprechrate als auch das Überleben zeigen sich bei Patienten mit Taxantherapie tendenziell günstiger, bei Patienten mit Radiochemotherapien sogar signifikant besser als die der Vergleichsbehandlungen. Bei der Dauer der Remissionen ergeben sich dagegen keine signifikanten Unterschiede. Stellt man das Hauptkriterium des Überlebens der Patienten mit fortgeschrittenem Bronchialkarzinom in Bezug zum Remissionsverhalten, so stellt man fest, dass Patienten deren Tumor auf die Therapie anspricht, signifikant länger leben. Bezieht man dagegen das Ansprechen mit unterschiedlichen Therapieansätzen auf das Überleben der Patienten, so zeigt sich bei Patienten mit Remission kein signifikanter Unterschied, ob sie mit taxanhaltigen oder taxanfreien Chemotherapien behandelt wurden. Demgegenüber leben Patienten, die Behandlungen ohne Strahlentherapie erhielten und Remission zeigen, signifikant länger als Responder mit Radiochemotherapien, obwohl die Ansprechrate der Radiochemotherapien signifikant besser ist und im gesamten Patientenkollektiv Patienten mit Radiochemotherapien signifikant länger leben als ohne Strahlentherapie. In Bezug auf die Therapieoptionen korreliert ein Ansprechen auf die Behandlung folglich nicht mit einer Verlängerung des Überlebens. Als bedeutende Vorhersagekriterien bezüglich des Überlebens haben sich das Tumorstadium bei Erstdiagnose wie auch der Allgemeinzustand der Patienten in der eigenen Studie bestätigt. Ein signifikant längeres Überleben und bessere Überlebensrate nach einem Jahr zeigen sowohl Patienten mit einem guten Allgemeinzustand, als auch Patienten im Stadium III, unabhängig vom Therapieansatz. Unabhängig vom Therapieansatz ist die Überlebenszeit der Patienten mit komplett abgeschlossener Therapie signifikant besser als die der abgebrochenen Behandlungsversuche. Auch die Ansprechrate ist tendenziell höher, keine Korrelation findet sich dagegen zu der Dauer des rezidivfreien Intervalls nach Remission. Zusammenfassend lässt sich sagen, dass Patienten mit inoperablem nicht-kleinzelligem Bronchialkarzinom in Bezug auf Therapiebelastung und Ansprechrate durch palliative Behandlungsansätze deutlich durch taxanhaltige Chemotherapien und Radiochemotherapien profitieren. Eine signifikante Verlängerung des Überlebens ist nur bei Radiochemotherapien verglichen mit Behandlungen ohne Radiatio zu beobachten. Dagegen zeigt sich beim Vergleich taxanhaltiger mit taxanfreien Chemotherapien kein signifikanter Unterschied der Überlebenszeiten, auch nach Aufteilung des Patientenkollektivs jeweils nach Stadien, Allgemeinzustand, Remissionsverhalten und Erhalt einer Kompletten Therapie oder Abbruch der Therapie. Signifikante Bedeutung für ein besseres Überleben haben aber allgemeine Prognosefaktoren, wie eine geringe Krankheitsausdehnung und ein guter Allgemeinzustand bei Therapiebeginn, wie auch ein Ansprechen auf die Therapie, allerdings unabhängig vom Behandlungsansatz. Auch andere Studien gehen davon aus, dass diese Faktoren letztlich auch die Dauer des Ansprechens und das Überleben bestimmen. Die meisten in dieser Analyse zitierten Studien weisen eine prinzipielle Effektivität der Polychemotherapien und kombinierten Radiochemotherapien auch beim nicht-kleinzelligen Bronchialkarzinom nach, sodass es heute keinesfalls gerechtfertigt ist, grundsätzlich auf eine Chemotherapie zu verzichten. Letztlich ist es aber nur in Studien möglich, durch Ueberprüfung und Weiterentwicklung alter und neuer Therapieansätze sowie Erprobung neuer Medikamente die Ergebnisse der Chemotherapie weiter zu verbessern.In this study we examined the effectivity of palliative therapystrategies for patients with non-small-cell lung cancer

Polyphase metamorphism and deformation in the Kalak and Reisa Nappe Complexes: unravelling the complex polyphase pre-Caledonian and Caledonian evolution of continental crust in northern Norway

International audienceThe Kalak and the Reisa Nappe Complexes (KNC and RNC) represent large parts of the Caledonides in northern Norway and allow the reconstruction of the pre- and syn-collisional history of the mid- to lower-crustal rocks of the orogen. We use a combination of field mapping, tectonostratigraphic description, phase equilibrium modelling and age dating to discuss their original relationships and subsequent evolution. The structurally lower KNC occurs as several nappes composed of orthogneisses and thick sequences of psammitic and pelitic metasedimentary gneisses with minor felsic and mafic intrusions. The structurally higher RNC includes three nappes; the Vaddas, Kåfjord and Nordmannvik Nappes. All three nappes are comprised of metasedimentary rocks (metapelites and metapsammites) and are intruded by gabbroic bodies. Caledonian metamorphism is recorded by a pervasive fabric throughout both nappe complexes. It increases in grade upwards from greenschist facies at the base of the KNC to amphibolite facies at the KNC-RNC boundary and to granulite facies in the Nordmannvik Nappe. In the KNC, the upper and lower parts record different pre-Caledonian histories. Age dating indicates that the lower part was affected by an event at c. 980-960 Ma, whereas the upper part experienced events at c. 850-820 Ma, c. 710 Ma and c. 580-520 Ma (Seiland Igneous Province). The upper nappes display pre-Caledonian amphibolite to granulite facies foliation (c. 710 Ma in age), while the lower nappes record upper greenschist-low amphibolite facies metamorphism associated with a different pre-Caledonian foliation . The difference in metamorphic conditions indicates that the upper and lower KNC represent different sections of basement juxtaposed by thrusting during Caledonian continental collision. The RNC rocks preserve a younger evolution. The Nordmannvik Nappe metasediments may have been deposited in the late Neoproterozoic, and are probably related to the Narvik Nappe, whereas the Vaddas and Kåfjord rocks were deposited in the late Ordovician-early Silurian. All three nappes record an anticlockwise P-T path, with high T, medium P metamorphism and mafic magmatism at ∼440 Ma, followed by higher P, lower T solid state shearing during nappe stacking at ∼430 Ma. Conditions at 440 Ma indicate that the migmatized Nordmannvik Nappe was deeper than the Kåfjord and Vaddas metasediments. The tectonic setting for the anticlockwise evolution is best described by magmatic underplating and heating of the three nappes in the subduction back-arc domain, which was quickly followed by an onset of subduction with subsequent nappe stacking and the RNC rocks probably formed parts of a back-arc during Ordovician-early Silurian subduction of the Seve Nappe rocks, outboard of the extended margin. Subduction rapidly followed the heating of these units caused by magmatic underplating (during back arc extension). The hot and weak crust was detached from the continental lithosphere and incorporated in the nappe their stacking during continental collision

Polyphase metamorphism and deformation in the Kalak and Reisa Nappe Complexes: unravelling the complex polyphase pre-Caledonian and Caledonian evolution of continental crust in northern Norway

International audienceThe Kalak and the Reisa Nappe Complexes (KNC and RNC) represent large parts of the Caledonides in northern Norway and allow the reconstruction of the pre- and syn-collisional history of the mid- to lower-crustal rocks of the orogen. We use a combination of field mapping, tectonostratigraphic description, phase equilibrium modelling and age dating to discuss their original relationships and subsequent evolution. The structurally lower KNC occurs as several nappes composed of orthogneisses and thick sequences of psammitic and pelitic metasedimentary gneisses with minor felsic and mafic intrusions. The structurally higher RNC includes three nappes; the Vaddas, Kåfjord and Nordmannvik Nappes. All three nappes are comprised of metasedimentary rocks (metapelites and metapsammites) and are intruded by gabbroic bodies. Caledonian metamorphism is recorded by a pervasive fabric throughout both nappe complexes. It increases in grade upwards from greenschist facies at the base of the KNC to amphibolite facies at the KNC-RNC boundary and to granulite facies in the Nordmannvik Nappe. In the KNC, the upper and lower parts record different pre-Caledonian histories. Age dating indicates that the lower part was affected by an event at c. 980-960 Ma, whereas the upper part experienced events at c. 850-820 Ma, c. 710 Ma and c. 580-520 Ma (Seiland Igneous Province). The upper nappes display pre-Caledonian amphibolite to granulite facies foliation (c. 710 Ma in age), while the lower nappes record upper greenschist-low amphibolite facies metamorphism associated with a different pre-Caledonian foliation . The difference in metamorphic conditions indicates that the upper and lower KNC represent different sections of basement juxtaposed by thrusting during Caledonian continental collision. The RNC rocks preserve a younger evolution. The Nordmannvik Nappe metasediments may have been deposited in the late Neoproterozoic, and are probably related to the Narvik Nappe, whereas the Vaddas and Kåfjord rocks were deposited in the late Ordovician-early Silurian. All three nappes record an anticlockwise P-T path, with high T, medium P metamorphism and mafic magmatism at ∼440 Ma, followed by higher P, lower T solid state shearing during nappe stacking at ∼430 Ma. Conditions at 440 Ma indicate that the migmatized Nordmannvik Nappe was deeper than the Kåfjord and Vaddas metasediments. The tectonic setting for the anticlockwise evolution is best described by magmatic underplating and heating of the three nappes in the subduction back-arc domain, which was quickly followed by an onset of subduction with subsequent nappe stacking and the RNC rocks probably formed parts of a back-arc during Ordovician-early Silurian subduction of the Seve Nappe rocks, outboard of the extended margin. Subduction rapidly followed the heating of these units caused by magmatic underplating (during back arc extension). The hot and weak crust was detached from the continental lithosphere and incorporated in the nappe their stacking during continental collision

Acute Toxicity Associated With the Recreational Use of the Novel Psychoactive Benzofuran N-methyl-5-(2 aminopropyl)benzofuran

N-methyl-5-(2 aminopropyl)benzofuran (5-MAPB) is a novel psychoactive benzofuran, created by N-methylation of 5-(2-aminopropyl)benzofuran (5-APB), which shares structural features with methylenedioxymethamphetamine (MDMA). To our knowledge, no case of 5-MAPB-related toxicity has been published in the scientific literature. We report a case of oral 5-MAPB exposure confirmed by liquid chromatography-tandem mass spectrometry in a 24-year-old previously healthy white man. Observed symptoms and signs such as paleness, cold and clammy skin, hypertension, elevated high-sensitive troponin T level, tachycardia, ECG change, diaphoresis, mild hyperthermia, mydriasis, tremor, hyperreflexia, clonus, agitation, disorientation, hallucinations, convulsions, reduced level of consciousness, and creatine kinase level elevation (305 IU/L) were compatible with undesired effects related to 5-APB or MDMA exposure. Signs and symptoms resolved substantially within 14 hours with aggressive symptomatic treatment, including sedation with benzodiazepines, external cooling, analgesia and sedation with fentanyl-propofol, and treatment with urapidil, an α-receptor-blocking agent. 5-MAPB showed first-order elimination kinetics with a half-life of 6.5 hours, comparable to the half-life of MDMA. According to the chemical structure, this case report, and users' Web reports, 5-MAPB appears to have an acute toxicity profile similar to that of 5-APB and MDMA, with marked vasoconstrictor effect

Gene augmentation of <i>LCA5</i>-associated Leber congenital amaurosis ameliorates bulge region defects of the photoreceptor ciliary axoneme

Leber congenital amaurosis (LCA) is a group of inherited retinal diseases characterized by early-onset, rapid loss of photoreceptor cells. Despite the discovery of a growing number of genes associated with this disease, the molecular mechanisms of photoreceptor cell degeneration of most LCA subtypes remain poorly understood. Here, using retina-specific affinity proteomics combined with ultrastructure expansion microscopy, we reveal the structural and molecular defects underlying LCA type 5 (LCA5) with nanoscale resolution. We show that LCA5-encoded lebercilin, together with retinitis pigmentosa 1 protein (RP1) and the intraflagellar transport (IFT) proteins IFT81 and IFT88, localized at the bulge region of the photoreceptor outer segment (OS), a region crucial for OS membrane disc formation. Next, we demonstrate that mutant mice deficient in lebercilin exhibited early axonemal defects at the bulge region and the distal OS, accompanied by reduced levels of RP1 and IFT proteins, affecting membrane disc formation and presumably leading to photoreceptor death. Finally, adeno-associated virus–based LCA5 gene augmentation partially restored the bulge region, preserved OS axoneme structure and membrane disc formation, and resulted in photoreceptor cell survival. Our approach thus provides a next level of assessment of retinal (gene) therapy efficacy at the molecular level.</p