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Fibrin glue repair leads to enhanced axonal elongation during early peripheral nerve regeneration in an in vivo mouse model

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Neural Regeneration Research 2015年第7期10卷 1166-1171页

作者： Georgios Koulaxouzidis Gernot Reim christian witzel Department of Plastic and Hand Surgery University of Freiburg Medical Centre Plastic and Reconstructive Surgery – Interdisciplinary Breast Center Charité–Universit?tsmedizin Berlin

Microsurgical suturing is the gold standard of nerve coaptation. Although literature on the usefulness of fibrin glue as an alternative is becoming increasingly available, it remains contradictory. Furthermore, no data exist on how both repair methods might influence the morphological aspects（arborization; branching） of early peripheral nerve regeneration. We used the sciatic nerve transplantation model in thy-1 yellow fluorescent protein mice（YFP; n = 10）. Pieces of nerve（1cm） were grafted from YFP-negative mice（n = 10） into those expressing YFP. We performed microsuture coaptations on one side and used fibrin glue for repair on the contralateral side. Seven days after grafting, the regeneration distance, the percentage of regenerating and arborizing axons, the number of branches per axon, the coaptation failure rate, the gap size at the repair site and the time needed for surgical repair were all investigated. Fibrin glue repair resulted in regenerating axons travelling further into the distal nerve. It also increased the percentage of arborizing axons. No coaptation failure was detected. Gap sizes were comparable in both groups. Fibrin glue significantly reduced surgical repair time. The increase in regeneration distance, even after the short period of time, is in line with the results of others that showed faster axonal regeneration after fibrin glue repair. The increase in arborizing axons could be another explanation for better functional and electrophysiological results after fibrin glue repair. Fibrin glue nerve coaptation seems to be a promising alternative to microsuture repair.

关键词： nerve regeneration fibrin glue peripheral nerve regeneration thy-1-YFP mice sciatic nerve branching arborisation neural regeneration

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N-Propionylmannosamine:using biochemical glycoengineering to promote peripheral nerve regeneration

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Neural Regeneration Research 2015年第12期10卷 1947-1948页

作者： Georgios Koulaxouzidis Werner Reutter christian witzel Department of Plastic and Hand Surgery University of Freiburg Medical Center Institut für Laboratoriumsmedizin Klinische Chemie und Pathobiochemie Charité-Universit?tsmedizin (Freie Universit?t Berlin) Plastic and Reconstructive Surgery Charité-Universit?tsmedizin Berlin

The peripheral nervous system,in contrast to the central nervous system,is capable of spontaneous ***,nerve reconstruction in the peripheral nervous system remains a major challenge,as the functional outcomes followin... 详细信息

关键词： N-Propionylmannosamine Figure Man ST UDP acid

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N-Propionylmannosamine stimulates axonal elongation in a murine model of sciatic nerve injury

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Neural Regeneration Research 2015年第6期10卷 976-981页

作者： christian witzel Werner Reutter G.Bj?rn Stark Georgios Koulaxouzidis Plastic and Reconstructive Surgery Interdisciplinary Breast CenterCharité–Universit?tsmedizin Berlin Institute of Laboratory Medicine Clinical Chemistry and PathobiochemistryCharité–Universit?tsmedizin Berlin Department of Plastic and Hand Surgery University of Freiburg Medical Centre

Increasing evidence indicates that sialic acid plays an important role during nerve regeneration. Sialic acids can be modified in vitro as well as in vivo using metabolic oligosaccharide engineering of the N-awl side chain. N-Propionylmannosamine （ManNProp） increases neurite outgrowth and accelerates the reestablishment of functional synapses in vitro. We investigated the influence of systemic ManNProp application using a specific in vivo mouse model. Using mice expressing axonal fluorescent proteins, we quantified the extension of regenerating axons, the number of regenerating axons, the number of arborising axons and the number of branches per axon 5 days after injury. Sciatic nerves from non-expressing mice were grafted into those expressing yellow fluorescent protein. We began a twice-daily intraperitoneal application of either peracetylated ManNProp （200 mg/kg） or saline solution 5 days before injury, and continued it until nerve harvest （5 days after transection）. ManNProp significantly increased the mean distance of axonal regeneration （2.49 mm vs. 1.53 mm; P 〈 0.005） and the number of arborizing axons （21% vs. 16%; P = 0.008） 5 days after sciatic nerve grafting. ManNProp did not affect the number of regenerating axons or the number of branches per arborizing axon. The biochemical glycoengineering of the N-acyl side chain of sialic acid might be a promising approach for improving peripheral nerve regeneration.

关键词： N-propionylmannosamine sialic acid glycoengineering sciatic nerve peripheral nerve branching arborisation Thyl-YFP mouse nerve regeneration

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A self-made, low-cost infrared system for evaluating the sciatic functional index in mice

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Neural Regeneration Research 2016年第5期11卷 829-834页

作者： Lukas Fricker Vincenzo Penna Florian Lampert G.Bjoern Stark christian witzel Georgios Koulaxouzidis Department of Plastic and Hand Surgery University of Freiburg Medical Center Plastic and Reconstructive Surgery Charité – Universit?tsmedizin Berlin

The sciatic functional index（SFI） is a popular parameter for peripheral nerve evaluation that relies on footprints obtained with ink and paper. Drawbacks include smearing artefacts and a lack of dynamic information during measurement. Modern applications use digitized systems that can deliver results with less analytical effort and fewer mice. However, the systems are expensive（€40,000）. This study aimed to evaluate the applicability and precision of a self-made, low-cost infrared system for evaluating SFI in mice. Mice were subjected to unilateral sciatic nerve crush injury（crush group; n = 7） and sham operation（sham group; n = 4）. They were evaluated on the day before surgery, the 2nd, 4th and 6th days after injury, and then every day up to the 23rd day after injury. We compared two SFI evaluation methods, i.e., conventional inkand-paper SFI（C-SFI） and our infrared system（I-SFI）. Our apparatus visualized footprints with totally internally reflected infrared light（950 nm） and a camera that can only detect this wavelength. Additionally we performed an analysis with the ladder beam walking test（LBWT） as a reference test. I-SFI assessment reduced the standard deviation by about 33 percent, from 11.6 to 7.8, and cut the variance around the baseline to 21 percent. The system thus requires fewer measurement repetitions and fewer animals, and cuts the cost of keeping the animals. The apparatus cost €321 to build. Our results show that the process of obtaining the SFI can be made more precise via digitization with a self-made, low-cost infrared system.

关键词： nerve regeneration sciatic nerve crush frustrated total internal reflection infrared ladder beamwalking test error rate mouse peripheral nerve injury neural regeneration

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Salinity Stress in Roots of Contrasting BarleyGenotypes Reveals Time-Distinct and Genotype-Specific Patterns for Defined Proteins

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Molecular Plant 2014年第2期7卷 336-355页

作者： Katja witzel Andrea Matros Marc Strickert Stephanie Kaspar Manuela Peukert Karl H. Mühling Andreas Börner Hans-Peter Mock Leibniz Institute of Plant Genetics and Crop Plant Research Corrensstrasse 3 06466 Gatersleben Germany Present address: Leibniz Institute of Vegetable and Ornamental Crops Theodor-Echtermeyer-Weg 1 14979 Großbeeren Germany Present address: University of Siegen Media Systems Group Hölderlinstrasse 3 57068 Siegen Germany Present address: Bruker Daltonik GmbH Fahrenheitstrasse 4 28359 Bremen Germany Institute of Plant Nutrition and Soil Science Christian Albrechts University Hermann-Rodewald-Strasse 2 24118 Kiel Germany

Soil salinity is one of the most severe abiotic stress factors threatening agriculture worldwide. Hence,particular interest exists in unraveling mechanisms leading to salt tolerance and improved crop plant performance onsaline soils. Barley is considered to be one of the most salinity-tolerant crops, but varying levels of tolerance are wellcharacterized. A proteomic analysis of the roots of two contrasting cultivars （cv. Steptoe and cv. Morex） is *** plants were exposed to a period of 1, 4, 7, or 10 d at 0, 100, or 150mM NaCI. The root proteome was analyzedbased on two-dimensional gel electrophoresis. A number of cultivar-specific and salinity stress-responsive proteins wereidentified. Mass spectrometry-based identification was successful for 74 proteins, and a hierarchical clustering analysisgrouped these into five clusters based on similarity of expression profile. The rank product method was applied to sta-tistically access the early and late responses, and this delivered a number of new candidate proteins underlying salinitytolerance in barley. Among these were some germin-like proteins, some pathogenesis-related proteins, and numerousas-yet uncharacterized proteins. Notably, proteins involved in detoxification pathways and terpenoid biosynthesis weredetected as early responsive to salinity and may function as a means of modulating growth-regulating mechanisms andmembrane stability via fine tuning of phytohormone and secondary metabolism in the root.

关键词： barley genotypes mass spectrometry proteome analysis salinity stress two-dimensional gel electrophoresis

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