Studying the Patterning Mechanisms and Cell Fates during Limb Regeneration in Ambystoma mexicanum

We studied patterning mechanisms and cell fates during limb regeneration in the axolotl. 1) It is crucial to understand the earliest events of patterning. Since it is technically challenging to study early events, we established single cell PCR. This new tool will allow us to obtain novel insight into the initial steps of limb patterning. 2)We have examined the roles of different tissues regarding their fates and features of proximo- distal patterning. Our strategy was to transplant GFP+ skin, skeleton, muscle and Schwann cells from transgenic donors to limbs of wild type hosts, amputate through the graft and analyze fluorescent progeny combined with the use of molecular markers. Our results revealed that different subpopulations of blastema cells exist regarding two aspects. First, we found that progeny of skin and skeleton have some tissue specific memory since they did not give rise to muscle lineages. However, cells of the skin contributed to other mesenchymal tissues like cartilage or tendons, while the majority of skeleton- derived cells undergoes self- renewal. Second, we performed one cellular and two molecular assays to investigate what tissues generate cells that exhibit features of proximo- distal patterning. Both assays revealed that Schwann cell- derived progeny do not display such features while progeny of skin, skeleton and muscle did. Therefore, we conclude that the blastema is a heterogeneous mix of cells regarding tissue lineages and features of proximo- distal patterning

Heat shock enhancer element-mediated expression of ß-glucuronidase in arabidopsis thaliana

Ein künstlich hergestelltes hitzeinduzierbares genetisches Element wurde in Arabidposis mithilfe eines Reportergens getestet und die induzierte Enzymaktivität quantitativ gemessen. Dazu wurde das Element in ein geeignetes DNA Konstrukt eingefügt, dieses in die zu testenden Pflanzen transformiert. Von diesen wurden einzelne Teile einem itzeschock ausgesetzt und die induzierte Enzymaktivität flourometrisch bestimmt.The idealized HSE multimer from zebrafish has been introduced into a binary shuttle vector in front of a GUS reporter gene under control of a CaMV minimal promoter. To achieve this, the GUS gene under control of a CaMV minimal promoter from p221.9 Gre6 was used to replace the same gene on pBI101.1 (under full CaMV promoter) to obtain a reporter GUS gene genetically silent when not enhanced by an extrernal factor such as a heat shock element. For the final test construct, the heat shock element from pSGH2 was introduced directly before the minimal promoter of the reporter GUS gene. This construct was then transformed into Agrobacterium tumefaciens by electroporation. Cultures from positive transformants (screened by antibiotic selection and reverse ligation) were used for Arabidopsis floral dip to transfer the vector into Arabidopsis plants. The transgenic lines were then tested for their heat shock phenotype, using a quantitative assay to evaluate the magnitude of inductio

Low-melting manganese(II)-based ionic liquids: syntheses, structures, properties and influence of trace impurities

The synthesis of more than 10 new magnetic ionic liquids with [MnX4]2 anions, X = Cl, NCS, NCO, is presented. Detailed structural information through single-crystal X-ray di raction is given for (DMDIm)[Mn(NCS)4], (BnEt3N)2[Mn(NCS)4], and {(Ph3P)2N}2[Mn(NCO4)] 0.6H2O, respectively. All compounds consist of discrete anions and cations with tetrahedrally coordinated Mn(II) atoms. They show paramagnetic behavior as expected for spin-only systems. Melting points are found for several systems below 100 C classifying them as ionic liquids. Thermal properties are investigated using di erential scanning calorimetry (DSC) measurements. The physicochemical properties of density, dynamic viscosity, electrolytic conductivity, and surface tension were measured temperature-dependent of selected samples. These properties are discussed in comparison to similar Co containing systems. An increasing amount of bromide impurity is found to a ect the surface tension only up to 3.3%

Tissue regeneration in dentistry: can salamanders provide insight?

The ability to regenerate damaged tissues would be of tremendous benefit for medicine and dentistry. Unfortunately, humans are unable to regenerate tissues such teeth, fingers or to repair injured spinal cord. With an aging population, health problems are more prominent and dentistry is no exception as loss of bone tissue in the orofacial sphere from periodontal disease is on the rise. Humans can repair oral soft tissues exceptionally well, however hard tissues, like bone and teeth, are devoid of the ability to repair well or at all. Fortunately, Mother Nature has solved nearly every problem that we would like to solve for our own benefit and tissue regeneration is no exception. By studying animals that can regenerate, like Axolotls (Mexican salamander), we hope to find ways to stimulate regeneration in humans. We will discuss the role of the transforming growth factor beta cytokines as they are central to wound healing in humans and regeneration in Axolotls. We will also compare wound healing in humans (skin and oral mucosa) to Axolotl skin wound healing and limb regeneration. Finally, we will address the problem of bone regeneration and present results in salamanders which indicate that in order to regenerate bone you need to recruit non-bone cells. Fundamental research, such as the work being done in animals that can regenerate, offers insight to help understand why some treatments are successful while others fail when it comes to specific tissues such as bones

At low water activity alpha-chymotrypsin is more active in an ionic liquid than in non-ionic organic solvents

The kinetics of the alpha-chymotrypsin catalysed transesterification of N-acetyl-l-phenylalanine ethyl ester with 1-butanol and the competing hydrolysis were evaluated at fixed water activity in two ionic liquids and two non-ionic organic solvents. In most respects the four solvents behaved similarly. However, at a water activity of 0.33, higher catalytic activity was observed in the ionic liquid, 1-butyl-3-methylimidazolium bis[(trifluoromethyl)sulfonyl]amide, than in the other solvents, and at a(w)=0.11 catalysis was only observed in this solvent

Determination of Postprandial Glycemic Responses by Continuous Glucose Monitoring in a Real-World Setting

Background: Self-monitoring of blood glucose using capillary glucose testing (C) has a number of shortcomings compared to continuous glucose monitoring (CGM). We aimed to compare these two methods and used blood glucose measurements in venous blood (IV) as a reference. Postprandial blood glucose levels were measured after 50 g oral glucose load and after the consumption of a portion of different foods containing 50 g of carbohydrates. We also evaluated the associations between postprandial glucose responses and the clinical characteristics of the participants at the beginning of the study. Methods: 12 healthy volunteers (age: 36 ± 17 years, BMI: 24.9 ± 3.5 kg/m2) ate white bread (WB) and whole grain (WG) bread and drank a 50 g glucose drink as reference. Postprandial glucose responses were evaluated by CGM, IV and C blood glucose measurements. Incremental area under the curve (AUCi) of postprandial blood glucose was calculated for 1 h (AUCi 0-60) and 2 h (AUCi 0-120). Results: After the consumption of white bread and whole grain bread, the AUCi 0-60 min did not differ between CGM and IV or C. AUCi 0-120 min of CGM showed no difference compared to C. Correlation analyses revealed a positive association of age with glucose AUCi 0-120 (r = 0.768; P = 0.004) and WG AUCi 0-120 (r = 0.758; P = 0.004); fasting blood glucose correlated with WG AUCi 0-120 (r = 0.838; P < 0.001). Conclusion: Despite considerable inter-individual variability of postprandial glycemic responses, CGM evaluated postprandial glycemic excursions which had comparable results compared to standard blood glucose measurements under real-life conditions. Associations of AUCi 0-60 and AUCi 0-120 postprandial glucose response with age or fasting blood glucose could be shown