Wirkmechanismen des MAP-Kinase Inhibitors CNI-1493 auf mikrogliale BV-2 Zellen und primäre Mikroglia

Ein zentrales Merkmal der Demenz vom Alzheimertyp (AD) ist die Akkumulation von β-Amyloid-Oligomeren sowie die Ausbildung einer β-Amyloid-induzierten Entzündungsreaktion im Gehirn. β-Amyloid entsteht aus einem Vorläuferprotein, dem APP, welches durch Sekretasen prozessiert wird. Die Bildung und der Abbau dieser Oligomere stellen einen zentralen Ansatz in der Entwicklung neuer Medikamente für AD dar. Insbesondere wird versucht, die Sekretasenaktivität zu beeinflussen, den Abbau von β-Amyloid zu fördern, die Aggregation von β-Amyloid zu hemmen sowie die durch die Oligomere bedingte Toxizität zu beeinflussen. Darüber hinaus steht die Modulation der β-Amyloid-induzierten Inflammation, bei der es zu einer starken Aktivierung mikroglialer Zellen kommt, im Focus des Interesses. In diesem Zusammenhang konnte gezeigt werden, dass der entzündungshemmende p38 MAPK Hemmstoff CNI-1493 in einem transgenen Mausmodel der Demenz vom Alzheimertyp zu einer deutlichen Reduktion der intrazerebralen β-Amyloid-Konzentration und gleichzeitig zu einer Verbesserung der Kognition dieser Tiere geführt hat. Allerdings sind die Wirk-mechanismen dieser Substanz nicht vollständig geklärt. Auch die besondere Wirkung auf die Mikroglia, einer zentralen Zelle bei der Erkrankung, ist noch unklar. Daher war das Ziel der vorliegenden Arbeit, zum einem die Interaktion zwischen CNI-1493 und β-Amyloid zu charakterisieren. Weiterhin wurde in vitro an primären Mikrogliazellen der Einfluss von CNI-1493 auf die Zytokinsekretion, die Aktivität der Sekretasen, die Toxizität von β-Amyloid und die Phagozytosefähigkeit untersucht. Primäre Mikroglia wurden aus Mesokortizes von Mausembryonen isoliert. Zusätzlich erfolgten die ersten Experimente an einer Zellkulturlinie. Zur Untersuchung der Zytotoxizität wurden MTT-Assays durchgeführt. Die Sekretion von Zytokinen und Amyloid-β wurde mit Hilfe von ELISAs untersucht. Die Fähigkeit der Zellen zur Phagozytose wurde durch FACS-Analysen überprüft. Die Interaktion zwischen CNI-1493 und β-Amyloid-Oligomeren wurde anhand von Westernblot bzw. Dot-Blot-Verfahren getestet. Die Aktivität der Sekretasen wurde indirekt durch RT-PCR dargestellt, während deren Funktion mit Hilfe von fluoro-metrischen Assays bestimmt wurde. Wir konnten zeigen, dass CNI-1493 in der Lage ist, die Bildung von toxischen Aggregations-formen des β-Amyloids zu hemmen. Dadurch ließ sich die β-Amyloid-induzierte Schädigung mikroglialer Zellen reduzieren. Auch die Sekretion von proinflammatorischen Zytokinen konnte durch CNI-1493 vermindert werden. Darüber hinaus ließ sich durch die Inkubation mit CNI-1493 die Phagozytose von β-Amyloid verbessern. Wir konnten auch zeigen, dass CNI-1493 die Funktion der α-Sekretase moduliert und dadurch die Induktion des nicht-amyloidogenen Weges der APP-Prozessierung induziert. In der vorliegenden Studie konnten wir relevante Wirkmechanismen von CNI-1493 in der mikroglialen Zellkultur aufklären. Unsere Daten zeigen, dass es nicht nur zu einem direkten Effekt von CNI-1493 auf Bildung toxischer β-Amyloid-Oligomere kommt, sondern über eine Inhibition des p38 MAPK Signalweges auch zur Modulation immunologischer Funktionen von mikroglialen Zellen. Über eine Aktivierung der α-Sekretase durch CNI-1493 ließ sich zudem eine Reduktion der β-Amyloid-Sekretion mikroglialer Zellen induzieren. In der Zusammen-schau dieser Ergebnisse scheint diese Substanz gerade aufgrund der unterschiedlichen Wirk-mechanismen für eine Therapie der AD geeignet. Berücksichtigt man die Tatsache, dass CNI-1493 bereits erfolgreich in einer klinischen Studie bei M. Crohn getestet wurde, wäre der Einsatz in einer klinischen Studie bei Patienten mit AD möglich

Placement and source effects of phosphate fertilizers on phosphorus availability and reaction products in two reduced-till soils: A greenhouse study

Citation: Khatiwada, Raju, et al. “Placement and Source Effects of Phosphate Fertilizers on Phosphorus Availability and Reaction Products in Two Reduced-Till Soils: A Greenhouse Study.” Soil Science, vol. 179, no. 3, Mar. 2014, pp. 141–152. journals.lww.com, doi:10.1097/SS.0000000000000055.Crop yields are limited primarily by unavailability of nutrients in agricultural soil. Adoption of reduced tillage and no-tillage systems leads to stratification of nutrients in surface soil, so management of phosphorus (P) in these systems is a major issue. The objective of this research was to understand the influence of placement (broadcast vs. deep-placed P) and fertilizer source (granular versus liquid P) on the reaction products of P under greenhouse conditions using soil columns. Phosphorus was added at a rate of 75 kg/ha to two soils: an acid soil from Manhattan, KS, and a slightly acid to neutral soil from Ottawa, KS. At 5 weeks after P application soil pH, resin-extractable P, and speciation of P in soils were determined at different distances from the point of fertilizer application. Scanning electron microscope with energy dispersive X-ray analysis(SEM-EDXA) and synchrotron-based X-ray absorption near-edge structure (XANES) spectroscopy were used to understand P speciation. Results for P fertilizer sources and placement with respect to resin-extractable P showed no clear differences between the treatments except for granular broadcast and granular deep-placed treatments in the Ottawa soil.Reaction products formed after application of P in two soils showed some clear differences. The XANES speciation revealed that Fe-P-like forms dominated in the acidic soil, whereas adsorbed and Fe and Ca-P-like forms dominated in the neutral to slightly acid soil. No clear trends in reaction products were detected with respect to P source or the P placement method. Furthermore, SEM-EDXA analysis of incubated fertilizer granules extracted from soils at 5 weeks showed enrichment of Al, Fe, and Ca in the zones of remaining P in incubated granules, indirectly indicating that these cations enter and or remaining in the granules and begin to react with P before the granules dissolve completely

The Mate Workbench - a tool for annotating XML corpora

This paper describes the design and implementation of the MATE workbench, a program which provides support for flexible display and editing of XML annotations, and complex querying of a set of linked files. The workbench was designed to support the annotation of XML coded linguistic corpora, but it could be used to annotate any kind of data, as it is not dependent on any particular annotation scheme. Rather than being a general purpose XMLaware editor it is a system for writing specialised editors tailored to a particular annotation task. A particular editor is defined using a transformation language, with suitable display formats and allowable editing operations. The workbench is written in Java, which means that it is platform-independent. This paper outlines the design of the workbench software and compares it with other annotation programs. 1. Introduction The annotation or markup of files with linguistic or other complex information usually requires either human coding or human ..

Crop Responses to AmiSorb in the North Central Region

Originally used to prevent scale in boilers, carpramid or thermal polyaspartate (copoly[(3-carboxypropionamide)( 2-carboxylmethyl) acetamide)] was brought to agriculture under the trade names AmiSorb and Magnet. It claimed to increase nutrient uptake through artificially increasing the volume of soil occupied by roots through increased root branching and root hair development. Under controlled hydroponic or greenhouse conditions, the use of carpramid increased nutrient uptake, some yield determining factors such as wheat tillering and in some cases, crop yield. Extensive field testing from 1996 to 1998 under various nutrient regimes, placements, forms, and timings resulted in very inconsistent performance. Averaged across all experiments for which data were available, small yield increases were observed for corn (+1.75 bushels/acre), soybean (+0.63 bushel/acre), wheat (+1.07 bushels/acre), and grain sorghum (+0.32 bushel/acre), but at best only about one-fourth of the experiments (27 percent for corn and wheat) showed statistically significant yield increases. Across all crops, only three experiments showed an economic advantage to using carpramid. An attempt was made to better define the conditions when responses were observed but no clear pattern emerged that would allow improved probability of predicting a positive response.https://lib.dr.iastate.edu/extension_pubs/1225/thumbnail.jp

Presenting signs and patient co-variables in Gaucher disease : outcome of the Gaucher Earlier Diagnosis Consensus (GED-C) Delphi initiative

© 2018 The Authors. Internal Medicine Journal by Wiley Publishing Asia Pty Ltd on behalf of Royal Australasian College of Physicians.Background: Gaucher disease (GD) presents with a range of signs and symptoms. Physicians can fail to recognise the early stages of GD owing to a lack of disease awareness, which can lead to significant diagnostic delays and sometimes irreversible but avoidable morbidities. Aim: The Gaucher Earlier Diagnosis Consensus (GED-C) initiative aimed to identify signs and co-variables considered most indicative of early type 1 and type 3 GD, to help non-specialists identify ‘at-risk’ patients who may benefit from diagnostic testing. Methods: An anonymous, three-round Delphi consensus process was deployed among a global panel of 22 specialists in GD (median experience 17.5 years, collectively managing almost 3000 patients). The rounds entailed data gathering, then importance ranking and establishment of consensus, using 5-point Likert scales and scoring thresholds defined a priori. Results: For type 1 disease, seven major signs (splenomegaly, thrombocytopenia, bone-related manifestations, anaemia, hyperferritinaemia, hepatomegaly and gammopathy) and two major co-variables (family history of GD and Ashkenazi-Jewish ancestry) were identified. For type 3 disease, nine major signs (splenomegaly, oculomotor disturbances, thrombocytopenia, epilepsy, anaemia, hepatomegaly, bone pain, motor disturbances and kyphosis) and one major co-variable (family history of GD) were identified. Lack of disease awareness, overlooking mild early signs and failure to consider GD as a diagnostic differential were considered major barriers to early diagnosis. Conclusion: The signs and co-variables identified in the GED-C initiative as potentially indicative of early GD will help to guide non-specialists and raise their index of suspicion in identifying patients potentially suitable for diagnostic testing for GD.Peer reviewedFinal Published versio

Speciation of phosphorus in a fertilized, reduced-till soil system: in-field treatment incubation study

Citation: Khatiwada, Raju, Ganga M. Hettiarachchi, David B. Mengel, and Mingwei Fei. “Speciation of Phosphorus in a Fertilized, Reduced-Till Soil System: In-Field Treatment Incubation Study.” Soil Science Society of America Journal 76, no. 6 (2012): 2006–18. https://doi.org/10.2136/sssaj2011.0299.Phosphorus management in reduced-tillage systems is a great concern for farmers. Conclusive positive results of deep-banding P fertilizers compared with broadcast application and the chemistry of reduced-tillage systems remain unclear. Knowledge of the dominant solid P species present in soil following application of P fertilizers and the resulting potential P availability would help us understand and efficiently manage P in reduced-tillage systems. The objective of this research was to study the influence of placement (broadcast vs. deep-band P), fertilizer source (granular vs. liquid P), and time on the reaction products of P under field conditions. Changes in soil pH, resin-extractable P, total P, and speciation of P were determined at different distances from the point of fertilizer application at 5 wk and 6 mo after P application at a rate of 75 kg ha−1 to a soil system that was under long-term reduced tillage. Resin-extractable P was lower for broadcast treatments compared with deep-band treatments for both time periods. Resin-extractable P was greater in the liquid P-treated soils than in the granular P-treated soils. Speciation results showed that granular P fertilizers tended to form Fe–P-like forms, whereas liquid forms remained in adsorbed P-like forms in the soil 5 wk after application; moreover, speciation results showed granular P fertilizers precipitated less when deep-banded. During the 6-mo period following application, reaction products of broadcast granular, broadcast liquid, and deep-band granular fertilizers transformed to Ca-phosphate or mixtures of Ca-, Fe- and adsorbed-phosphate-like forms, whereas deep-band liquid P remained as mainly adsorbed P-like forms. Deep-banding of P would most likely provide a solution that is both agronomically and environmentally efficient for reduced-till farmers