Green Synthesis of Glycopolymers Using an Enzymatic Approach

beta-Glucosidase and horseradish peroxidase (HRP) are used as biocatalysts in aqueous solution for the enzymatic synthesis of glycomonomers and the respective enzymatic polymerization toward glycopolymers. The biocatalytically synthesized monomers contain (meth)acrylate functionalities that are able to be polymerized by an enzyme-initiated polymerization using an HRP/hydrogen peroxide/acetylacetone ternary system. The structure of the glycomonomers and the respective glycopolymers as well as the monomer conversion after the reaction are determined by H-1 NMR spectroscopy. The synthesized glycopolymers have a dispersity and a number-average molecular weight up to 5.8 and 297 kg mol(-1), respectively. Thermal and degradation properties of the glycopolymers are studied by differential scanning calorimetry and thermogravimetric analysis. In addition, preparation of glycopolymers via conventional free radical polymerization is performed and the properties of the obtained polymers are compared with the enzymatically synthesized glycopolymers

Rangelands Vegetation Mapping at Species Composition Level Using the \u3cb\u3eSPiCla\u3c/b\u3e Method: \u3cb\u3eS\u3c/b\u3eDM Based \u3cb\u3ePi\u3c/b\u3exel \u3cb\u3eCla\u3c/b\u3essification and Fuzzy Accuracy. A New Approach of Map Making

Vegetation maps have been made since centuries. The vegetation cover was represented as homogeneous mapping units (polygons), representing different vegetation types, where each type consists a combination of different plant species (floristic composition). More recent, with the use of satellite imagery, the polygons have been replaced by pixels with similar content as the polygon maps. In both approaches, field-observations were linked to the mapping units (polygons or pixels) often resulting in a complex of different vegetation types per mapping unit. In our new approach field data (sample points) on presence and abundance of individual grass species are spatially extrapolated based on a set of environmental layers, using the species distribution modelling approach (SDM). When combined, each pixel will contain its own set of information about the vegetation structure and its floristic composition. This new methodology (SPiCla) results in a very accurate and detailed vegetation map at pixel level, allowing extraction of very detailed, accurate and easy to update spatial information on e.g., forage production and quality (palatability) for rangelands management. As no exact boundaries exist, but only gradients, we introduced fuzzy accuracy. The resolution mainly depends on the resolution of (or one of) the environmental layers used, scale of interest and workability. The methodology is generic and applicable to any other region in the world

Targeted Inhibition of miRNA Maturation with Morpholinos Reveals a Role for miR-375 in Pancreatic Islet Development

Several vertebrate microRNAs (miRNAs) have been implicated in cellular processes such as muscle differentiation, synapse function, and insulin secretion. In addition, analysis of Dicer null mutants has shown that miRNAs play a role in tissue morphogenesis. Nonetheless, only a few loss-of-function phenotypes for individual miRNAs have been described to date. Here, we introduce a quick and versatile method to interfere with miRNA function during zebrafish embryonic development. Morpholino oligonucleotides targeting the mature miRNA or the miRNA precursor specifically and temporally knock down miRNAs. Morpholinos can block processing of the primary miRNA (pri-miRNA) or the pre-miRNA, and they can inhibit the activity of the mature miRNA. We used this strategy to knock down 13 miRNAs conserved between zebrafish and mammals. For most miRNAs, this does not result in visible defects, but knockdown of miR-375 causes defects in the morphology of the pancreatic islet. Although the islet is still intact at 24 hours postfertilization, in later stages the islet cells become scattered. This phenotype can be recapitulated by independent control morpholinos targeting other sequences in the miR-375 precursor, excluding off-target effects as cause of the phenotype. The aberrant formation of the endocrine pancreas, caused by miR-375 knockdown, is one of the first loss-of-function phenotypes for an individual miRNA in vertebrate development. The miRNA knockdown strategy presented here will be widely used to unravel miRNA function in zebrafish

Differences in vertebrate microRNA expression

MicroRNAs (miRNAs) attenuate gene expression by means of translational inhibition and mRNA degradation. They are abundant, highly conserved, and predicted to regulate a large number of transcripts. Several hundred miRNA classes are known, and many are associated with cell proliferation and differentiation. Many exhibit tissue-specific expression, which aids in evaluating their functions, and it has been assumed that their high level of sequence conservation implies a high level of expression conservation. A limited amount of data supports this, although discrepancies do exist. By comparing the expression of ≈100 miRNAs in medaka and chicken with existing data for zebrafish and mouse, we conclude that the timing and location of miRNA expression is not strictly conserved. In some instances, differences in expression are associated with changes in miRNA copy number, genomic context, or both between species. Variation in miRNA expression is more pronounced the greater the differences in physiology, and it is enticing to speculate that changes in miRNA expression may play a role in shaping the physiological differences produced during animal development

Joubert syndrome: genotyping a Northern European patient cohort

Joubert syndrome (JBS) is a rare neurodevelopmental disorder belonging to the group of ciliary diseases. JBS is genetically heterogeneous, with >20 causative genes identified to date. A molecular diagnosis of JBS is essential for prediction of disease progression and genetic counseling. We developed a targeted next-generation sequencing (NGS) approach for parallel sequencing of 22 known JBS genes plus 599 additional ciliary genes. This method was used to genotype a cohort of 51 well-phenotyped Northern European JBS cases (in some of the cases, Sanger sequencing of individual JBS genes had been performed previously). Altogether, 21 of the 51 cases (41%) harbored biallelic pathogenic mutations in known JBS genes, including 14 mutations not previously described. Mutations in C5orf42 (12%), TMEM67 (10%), and AHI1 (8%) were the most prevalent. C5orf42 mutations result in a purely neurological Joubert phenotype, in one case associated with postaxial polydactyly. Our study represents a population-based cohort of JBS patients not enriched for consanguinity, providing insight into the relative importance of the different JBS genes in a Northern European population. Mutations in C5orf42 are relatively frequent (possibly due to a Dutch founder mutation) and mutations in CEP290 are underrepresented compared with international cohorts. Furthermore, we report a case with heterozygous mutations in CC2D2A and B9D1, a gene associated with the more severe Meckel–Gruber syndrome that was recently published as a potential new JBS gene, and discuss the significance of this finding

antiSMASH 6.0: improving cluster detection and comparison capabilities

Microbial Biotechnolog

Potato tuber pectin structure is influenced by pectin methyl esterase activity and impacts on cooked potato texture

Although cooked potato tuber texture is an important trait that influences consumer preference, a detailed understanding of tuber textural properties at the molecular level is lacking. Previous work has identified tuber pectin methyl esterase activity (PME) as a potential factor impacting on textural properties. In this study, tuber PME isoform and gene expression profiles have been determined in potato germplasm with differing textural properties as assessed using an amended wedge fracture method and a sloughing assay, revealing major differences between the potato types. Differences in pectin structure between potato types with different textural properties were revealed using monoclonal antibodies specific for different pectic epitopes. Chemical analysis of tuber pectin clearly demonstrated that, in tubers containing a higher level of total PME activity, there was a reduced degree of methylation of cell wall pectin and consistently higher peak force and work done values during the fracture of cooked tuber samples, demonstrating the link between PME activity, the degree of methylation of cell wall pectin, and cooked tuber textural properties