Computer-assisted and patient-specific 3-D planning and evaluation of a single-cut rotational osteotomy for complex long-bone deformities

Malunion after long bone fracture results in an incorrect position of the distal bone segment. This misalignment may lead to reduced function of the limb, early osteoarthritis and chronic pain. An established treatment option is a corrective osteotomy. For complex malunions, a single-cut rotational osteotomy is sometimes preferred in cases of angular deformity in three dimensions. However, planning and performing this type of osteotomy is relatively complex. This report describes a computer-assisted method for 3-D planning and realizing a single-cut rotational osteotomy with a patient-specific cutting guide for orienting the osteotomy and an angled jig for adjusting the rotation angle. The accuracy and reproducibility of the method is evaluated experimentally using plastic bones. In addition, complex rotational deformities are simulated by a computer to investigate the relation between deformity and correction parameters. The computed relation between deformity and correction parameters enables the surgeon to judge the feasibility of a single-cut rotational osteotomy. This appears possible for deformities combining axial misalignment with sufficient axial rotation. The proposed 3-D method of preoperative planning and transfer with a patient-specific cutting guide and angled jig renders the osteotomy procedure easily applicable, accurate, reproducible, and is a good alternative for complex and expensive navigation systems

Automated Three-Dimensional Detection and Shape Classification of Dendritic Spines from Fluorescence Microscopy Images

A fundamental challenge in understanding how dendritic spine morphology controls learning and memory has been quantifying three-dimensional (3D) spine shapes with sufficient precision to distinguish morphologic types, and sufficient throughput for robust statistical analysis. The necessity to analyze large volumetric data sets accurately, efficiently, and in true 3D has been a major bottleneck in deriving reliable relationships between altered neuronal function and changes in spine morphology. We introduce a novel system for automated detection, shape analysis and classification of dendritic spines from laser scanning microscopy (LSM) images that directly addresses these limitations. The system is more accurate, and at least an order of magnitude faster, than existing technologies. By operating fully in 3D the algorithm resolves spines that are undetectable with standard two-dimensional (2D) tools. Adaptive local thresholding, voxel clustering and Rayburst Sampling generate a profile of diameter estimates used to classify spines into morphologic types, while minimizing optical smear and quantization artifacts. The technique opens new horizons on the objective evaluation of spine changes with synaptic plasticity, normal development and aging, and with neurodegenerative disorders that impair cognitive function

Genome Characterization of the Oleaginous Fungus Mortierella alpina

Mortierella alpina is an oleaginous fungus which can produce lipids accounting for up to 50% of its dry weight in the form of triacylglycerols. It is used commercially for the production of arachidonic acid. Using a combination of high throughput sequencing and lipid profiling, we have assembled the M. alpina genome, mapped its lipogenesis pathway and determined its major lipid species. The 38.38 Mb M. alpina genome shows a high degree of gene duplications. Approximately 50% of its 12,796 gene models, and 60% of genes in the predicted lipogenesis pathway, belong to multigene families. Notably, M. alpina has 18 lipase genes, of which 11 contain the class 2 lipase domain and may share a similar function. M. alpina's fatty acid synthase is a single polypeptide containing all of the catalytic domains required for fatty acid synthesis from acetyl-CoA and malonyl-CoA, whereas in many fungi this enzyme is comprised of two polypeptides. Major lipids were profiled to confirm the products predicted in the lipogenesis pathway. M. alpina produces a complex mixture of glycerolipids, glycerophospholipids and sphingolipids. In contrast, only two major sterol lipids, desmosterol and 24(28)-methylene-cholesterol, were detected. Phylogenetic analysis based on genes involved in lipid metabolism suggests that oleaginous fungi may have acquired their lipogenic capacity during evolution after the divergence of Ascomycota, Basidiomycota, Chytridiomycota and Mucoromycota. Our study provides the first draft genome and comprehensive lipid profile for M. alpina, and lays the foundation for possible genetic engineering of M. alpina to produce higher levels and diverse contents of dietary lipids

Trichoderma longibrachiatum acetyl xylan esterase 1 enhances hemicellulolytic preparations to degrade corn silage polysaccharides

Supplementation of a Trichoderma longibrachiatum preparation to an industrial Aspergillus niger/Talaromyces emersonii enzyme mixture demonstrated synergy for the saccharification of corn silage water-unextractable solids (WUS). Sub-fractions of the crude T. longibrachiatum preparation obtained after chromatography were analyzed regarding their hydrolytic activity. An acetyl xylan esterase 1 [Axe1, carbohydrate esterase (CE) family 5]-enriched sub-fraction closely mimicked the hydrolytic gain as obtained by supplementation of the complete, crude enzyme mixture (increase of 50%, 62% and 29% for Xyl, Ara and Glc, respectively). The acetic acid released from model polysaccharides (WUS) and oligosaccharides [neutral (AcXOS) and acidic (AcUXOS) xylo-oligosaccharides] by Axe1 was two and up to six times higher compared to the acetic acid released by acetyl xylan esterase A (AxeA, CE 1). Characterization of Axe1 treated AcXOS and AcUXOS revealed deacetylation of oligosaccharides that were not deacetylated by AxeA or the A. niger/T. emersonii preparation

Acetyl Xylan Esterase Axe1 (T. reesei, Carbohydrate Esterase Family 5) Supplemented to a (Hemi)cellulolytic Preparation Enhances Degradation of Recalcitrant Corn Silage Polysaccharides

The increase in hydrolytic activity towards corn silage water unextractable solids by supplementation of acetyl xylan esterase 1 (T. reesei, TrAxe1), belonging to arbohydrate esterase (CE) family 5, to an A. niger / T. emersonii enzyme preparation is presented. TrAxe1 was cloned and expressed in A niger. The hydrolytic activity of TrAxe1 was analysed with a p-nitrophenyl-acetate standard assay and by the measurement of the acetic acid released from neutral and acidic eucalyptus xylo-oligosaccharides. The activity obtained for TrAxe1 was compared to acetyl xylan esterases belonging to CE families 1, 5 and 16 from A. niger. High activities were obtained for the CE5 classified acetyl xylan esterases from T. reesei and A. niger. The data obtained confirm the suggested increase in hydrolytic activity by supplementation of CE5 classified acetyl xylan esterases to the (hemi)cellulolytic A. niger / T. emersonii preparation (Neumüller et al. 2014b

Quantifying near metal visibility using dual energy computed tomography and iterative metal artifact reduction in a fracture phantom

Purpose: To quantitatively assess CT image quality and fracture visibility using virtual monochromatic imaging and iterative metal artifact reduction (iMAR) in a femoral bone fracture phantom with different fixation implants. Methods: A custom made phantom was scanned at 120-kVp and 140-kVp single-energy and 100/150-kVp dual-energy. Three stainless steel and two titanium implants with different thicknesses were placed on the phantom containing simulated one and two mm fractures. Single-energy CT images were reconstructed with and without iMAR, while DECT images were reconstructed at monochromatic energies between 70 and 190 keV. Non-metal scans were used as a reference. A Fourier power spectrum method and fracture model were used to analyze several anatomical areas. Results: CT-value deviations of titanium implants were much lower compared to stainless steel implants. These deviations decreased for both DECT and iMAR. Fracture visibility, measured with the fracture model, improved the most when DECT was used while artifact reduction benefitted more from iMAR. The optimal monochromatic energy for metal artifact reduction, based on CT-value deviation, varied for each metal between 130 and 150 keV. The fracture model provided a signal-to-noise ratio for the near metal fracture visibility, providing the optimal keV. Conclusion: iMAR and high keV monochromatic images extracted from DECT both reduce metal artifacts caused by different metal fixation implants. Quantitative femoral phantom results show that DECT is superior to iMAR regarding fracture visualization adjacent to metal fixation implants. The introduction of new artifacts when using iMAR impedes its value in near metal fixation implant imaging

Synergistic action of enzyme preparations towards recalcitrant corn silage polysaccharides

Corn silage, its water unextractable solids (WUS) and enzyme recalcitrant solids (ErCS) and an industrial corn silage-based anaerobic fermentation residue (AFR) represent corn substrates with different levels of recalcitrance. Compositional analysis reveals different levels of arabinoxylan substitution for WUS, ErCS and AFR, being most pronounced regarding acetic acid, glucuronic acid- and arabinose content. By screening for enzymatic degradation of WUS, ErCS and AFR, enzyme preparations exhibiting high conversion rates were identified. Furthermore significant synergistic effects were detected by blending Aspergillus niger/Talaromyces emersonii culture filtrates with various enzymes. These findings clearly highlight a necessity for a combinatorial use of enzyme preparations towards substrates with high recalcitrance characteristics to reach high degrees of degradation. Enzyme blends were identified, outperforming the individual commercial preparations. These enzyme preparations provide a basis for new, designed enzyme mixtures for corn polysaccharide degradation as a source of necessary, accessory enzyme activities

Positional preferences of acetyl esterases from different CE families towards acetylated 4-O-methyl glucuronic acid-substituted xylo-oligosaccharides

Background Acetylation of the xylan backbone restricts the hydrolysis of plant poly- and oligosaccharides by hemicellulolytic enzyme preparations to constituent monosaccharides. The positional preferences and deacetylation efficiencies of acetyl esterases from seven different carbohydrate esterase (CE) families towards different acetylated xylopyranosyl units (Xylp) - as present in 4-O-methyl-glucuronic acid (MeGlcA)-substituted xylo-oligosaccharides (AcUXOS) derived from Eucalyptus globulus - were monitored by 1H NMR, using common conditions for biofuel production (pH 5.0, 50°C). Results Differences were observed regarding the hydrolysis of 2-O, 3-O, and 2,3-di-O acetylated Xylp and 3-O acetylated Xylp 2-O substituted with MeGlcA. The acetyl esterases tested could be categorized in three groups having activities towards (i) 2-O and 3-O acetylated Xylp, (ii) 2-O, 3-O, and 2,3-di-O acetylated Xylp, and (iii) 2-O, 3-O, and 2,3-di-O acetylated Xylp, as well as 3-O acetylated Xylp 2-O substituted with MeGlcA at the non-reducing end. A high deacetylation efficiency of up to 83% was observed for CE5 and CE1 acetyl esterases. Positional preferences were observed towards 2,3-di-O acetylated Xylp (TeCE1, AnCE5, and OsCE6) or 3-O acetylated Xylp (CtCE4). Conclusions Different positional preferences, deacetylation efficiencies, and initial deacetylation rates towards 2-O, 3-O, and 2,3-di-O acetylated Xylp and 3-O acetylated Xylp 2-O substituted with MeGlcA were demonstrated for acetyl esterases from different CE families at pH 5.0 and 50°C. The data allow the design of optimal, deacetylating hemicellulolytic enzyme mixtures for the hydrolysis of non-alkaline-pretreated bioenergy feedstocks