Lignan Derivatives from Krameria lappacea Roots Inhibit Acute Inflammation in Vivo and Pro-inflammatory Mediators in Vitro

The roots of Krameria lappacea are used traditionally against oropharyngeal inflammation. So far, the astringent and antimicrobial properties of its proanthocyanidin constituents are considered to account for the anti-inflammatory effect. The aim of the present study was to characterize pharmacologically a lipophilic extract of K. lappacea roots and several isolated lignan derivatives (111) in terms of their putative anti-inflammatory activity. The dichloromethane extract (ID50 77 \u3bcg/cm2) as well compounds 111 (ID50 0.310.60 \u3bcmol/cm2) exhibited topical antiedematous properties comparable to those of indomethacin (ID50 0.29 \u3bcmol/cm2) in a mouse ear in vivo model. Two of the most potent compounds, 2-(2-hydroxy-4-methoxyphenyl)-5-(3-hydroxypropyl)benzofuran (5) and (+)-conocarpan (7), were studied regarding their time-dependent edema development and leukocyte infiltration up to 48 h after croton oil-induced dermatitis induction, and they showed activity profiles similar to that of hydrocortisone. In vitro studies of the isolated lignan derivatives demonstrated the inhibition of NFkB, cyclooxygenase-1 and -2, 5-lipoxygenase, and microsomal prostaglandin E2 synthase-1 as well as antioxidant properties, as mechanisms possibly contributing to the observed in vivo effects. The present findings not only support the ethnopharmacological use of K. lappacea roots but also reveal that the isolated lignan derivatives contribute strongly to the anti-inflammatory activity of this herbal drug

Minimal information for studies of extracellular vesicles 2018 (MISEV2018):a position statement of the International Society for Extracellular Vesicles and update of the MISEV2014 guidelines

The last decade has seen a sharp increase in the number of scientific publications describing physiological and pathological functions of extracellular vesicles (EVs), a collective term covering various subtypes of cell-released, membranous structures, called exosomes, microvesicles, microparticles, ectosomes, oncosomes, apoptotic bodies, and many other names. However, specific issues arise when working with these entities, whose size and amount often make them difficult to obtain as relatively pure preparations, and to characterize properly. The International Society for Extracellular Vesicles (ISEV) proposed Minimal Information for Studies of Extracellular Vesicles (“MISEV”) guidelines for the field in 2014. We now update these “MISEV2014” guidelines based on evolution of the collective knowledge in the last four years. An important point to consider is that ascribing a specific function to EVs in general, or to subtypes of EVs, requires reporting of specific information beyond mere description of function in a crude, potentially contaminated, and heterogeneous preparation. For example, claims that exosomes are endowed with exquisite and specific activities remain difficult to support experimentally, given our still limited knowledge of their specific molecular machineries of biogenesis and release, as compared with other biophysically similar EVs. The MISEV2018 guidelines include tables and outlines of suggested protocols and steps to follow to document specific EV-associated functional activities. Finally, a checklist is provided with summaries of key points

Vorbereitungsstand zur EU Verordnung 536/2014 in den Mitgliedstaaten

The upcoming Regulation EU 536/2014 for clinical trials of medicinal products for human use requires multinational cooperation in the assessment of clinical trial applications by the member states concerned as well as one single decision per member state concerned, supported by the new EU Portal and database system. The implementation makes national reorganisation of the processes necessary, especially coordination and cooperation between the national competent authorities and the ethics committees, necessary. A brief overview of implementation status with regard to national law adaptations, (re-) organisation, information technology systems as well as national or multinational pilot projects in 7 of 28 member states is given. Within these member states some national laws have been adapted already, while others will be soon. The national reorganisation covers mostly administrative organisation and coordination especially within the ethics committees. Overall, it is foreseen that a reduced number of ethics committees will be involved in the approval of clinical trials on medicinal products within the member states. Most of the seven member states expect that in addition to the new EU Portal/database system a national IT system for national cooperation and interaction will be necessary. Therefore, an interface within the EU system for national systems is essential. In order to test the new processes some member states are running national pilot projects or are planning them. In addition, almost all participate in the voluntary multinational assessment of clinical trials on medicinal products, which had existed since 2009, a few also in cooperation with their ethics committees. The member states are confident that all national processes will be in place when the EU regulation becomes applicable.Die kommende Verordnung EU 536/2014 über klinische Prüfungen mit Humanarzneimitteln sieht neben einer gemeinsamen Bewertung der betroffenen Mitgliedstaaten nur noch eine Entscheidung zur Genehmigung einer klinischen Prüfung von Arzneimitteln je betroffenem Mitgliedstaat vor, unterstützt durch das neue EU-Portal/Datenbanksystem. Im Rahmen der Implementierung werden nationale Reorganisationen im Verfahren notwendig, insbesondere zur Koordination und Kooperation zwischen nationaler Behörde und Ethikkommission/en. Es wird ein kurzer Überblick des nationalen Vorbereitungsstands gegeben bezüglich der Gesetzgebung, der (Re‑)Organisation, der Informationstechnik-Systeme sowie nationaler oder multinationaler Pilotprojekte in sieben von 28 Mitgliedstaaten. In diesen Mitgliedstaaten sind einige Gesetze schon angepasst oder stehen kurz davor. Die nationale Reorganisation betrifft zumeist die administrative Organisation und Koordination, insbesondere bei den Ethikkommissionen. Insgesamt ist abzusehen, dass sich weniger Ethikkommissionen als bisher an dem Genehmigungsverfahren zur klinischen Prüfung von Arzneimitteln in den Mitgliedstaaten beteiligen werden. Die meisten der sieben Mitgliedstaaten gehen von der Notwendigkeit eines nationalen IT-Systems für die nationale Koordination und Interaktion aus, zusätzlich zum neuen EU-Portal/Datenbanksystem. Folglich ist eine Schnittstelle im EU-System zu den nationalen Systemen essenziell. Zur Testung der neuen Prozesse laufen in einigen Mitgliedstaaten nationale Pilotprojekte, andere sind geplant. Zusätzlich beteiligen sich fast alle an der seit dem Jahre 2009 existierenden freiwilligen multinationalen Bewertung klinischer Prüfungen von Arzneimitteln; einige auch in Zusammenarbeit mit ihren Ethikkommissionen. Die Mitgliedstaaten sind zuversichtlich, dass zur Anwendung der EU-Verordnung alle nationalen Prozesse verfügbar sind

Engineering of Promoter Replacement Cassettes for Fine-Tuning of Gene Expression in Saccharomyces cerevisiae

The strong overexpression or complete deletion of a gene gives only limited information about its control over a certain phenotype or pathway. Gene function studies based on these methods are therefore incomplete. To effect facile manipulation of gene expression across a full continuum of possible expression levels, we recently created a library of mutant promoters. Here, we provide the detailed characterization of our yeast promoter collection comprising 11 mutants of the strong constitutive Saccharomyces cerevisiae TEF1 promoter. The activities of the mutant promoters range between about 8% and 120% of the activity of the unmutated TEF1 promoter. The differences in reporter gene expression in the 11 mutants were independent of the carbon source used, and real-time PCR confirmed that these differences were due to varying levels of transcription (i.e., caused by varying promoter strengths). In addition to a CEN/ARS plasmid-based promoter collection, we also created promoter replacement cassettes. They enable genomic integration of our mutant promoter collection upstream of any given yeast gene, allowing detailed genotype-phenotype characterizations. To illustrate the utility of the method, the GPD1 promoter of S. cerevisiae was replaced by five TEF1 promoter mutants of different strengths, which allowed analysis of the impact of glycerol 3-phosphate dehydrogenase activity on the glycerol yield