Relaxometry and brain myelin quantification with synthetic MRI in MS subtypes and their associations with spinal cord atrophy.

Immune-mediated demyelination and neurodegeneration are pathophysiological hallmarks of Multiple Sclerosis (MS) and main drivers of disease related disability. The principal method for evaluating qualitatively demyelinating events in the clinical context is contrast-weighted magnetic resonance imaging (MRI). Moreover, advanced MRI sequences provide reliable quantification of brain myelin offering new opportunities to study tissue pathology in vivo. Towards neurodegenerative aspects of the disease, spinal cord atrophy - besides brain atrophy - is a powerful and validated predictor of disease progression. The etiology of spinal cord volume loss is still a matter of research, as it remains unclear whether the impact of local lesion pathology or the interaction with supra- and infratentorial axonal degeneration and demyelination of the long descending and ascending fiber tracts are the determining factors. Quantitative synthetic MR using a multiecho acquisition of saturation recovery pulse sequence provides fast automatic brain tissue and myelin volumetry based on R1 and R2 relaxation rates and proton density quantification, making it a promising modality for application in the clinical routine. In this cross sectional study a total of 91 MS patients and 31 control subjects were included to investigate group differences of global and regional measures of brain myelin and relaxation rates, in different MS subtypes, using QRAPMASTER sequence and SyMRI postprocessing software. Furthermore, we examined associations between these quantitative brain parameters and spinal cord atrophy to draw conclusions about possible pathophysiological relationships. Intracranial myelin volume fraction of the global brain exhibited statistically significant differences between control subjects (10.4%) and MS patients (RRMS 9.4%, PMS 8.1%). In a LASSO regression analysis with total brain lesion load, intracranial myelin volume fraction and brain parenchymal fraction, the intracranial myelin volume fraction was the variable with the highest impact on spinal cord atrophy (standardized coefficient 4.52). Regional supratentorial MRI metrics showed altered average myelin volume fraction, R1, R2 and proton density in MS patients compared to controls most pronounced in PMS. Interestingly, quantitative MRI parameters in supratentorial regions showed strong associations with upper cord atrophy, suggesting an important role of brain diffuse demyelination on spinal cord pathology possibly in the context of global disease activity. R1, R2 or proton density of the thalamus, cerebellum and brainstem correlated with upper cervical cord atrophy, probably reflecting the direct functional connection between these brain structures and the spinal cord as well as the effects of retrograde and anterograde axonal degeneration. By using Synthetic MR-derived myelin volume fraction, we were able to effectively detect significant differences of myelination in relapsing and progressive MS subtypes. Total intracranial brain myelin volume fraction seemed to predict spinal cord volume loss better than brain atrophy or total lesion load. Furthermore, demyelination in highly myelinated supratentorial regions, as an indicator of diffuse disease activity, as well as alterations of relaxation parameters in adjacent infratentorial and midbrain areas were strongly associated with upper cervical cord atrophy.

In Vitro Ion Release of Wires in Removable Orthodontic Appliances.

Various orthodontic wire compositions and configurations are present on the market for removable appliances; however, there have still been only few studies focusing on the effect of resin color and additives such as glitter on corrosion of metallic wires under different conditions. Thus, the aim of the study was to compare concentrations of released ions (aluminium, chromium, nickel) in a corrosive medium under three different conditions: non-loaded wires, loaded wires, and non-loaded wires treated with Kukis® cleaning tablets. Six different wires made of three types of steel alloy were embedded in PMMA resin leaving one centimetre of each wire emerging from the resin to come into contact with the corrosive medium. Glitter particles were added to half of the produced test specimens. For the unloaded test series, five specimens of each group were covered in a petri dish with 50 mL of corrosive medium (pH 2.3) following EN-ISO 10271 for seven days at 37 °C. The wires for the mechanically loaded test specimens overlapped the resin by 5 cm and were clamped into a time-switched electric drive for a defined period of time before the samples were taken after a testing time of 7 days. In the third group, unloaded test specimens were transferred from their petri dishes into the prepared Kukis® solution every 24 h before being stored in the corrosive medium. Inductively coupled plasma mass spectrometry (ICP-MS) was used to quantify the specific ions in the corrosive solution. Statistical analysis showed that the mechanical loading of all wires could significantly raise the diffusion of ions into the corrosive medium. The colour of the resin did not affect the concentration of the released ions. The Kukis® cleaning tabs could not lower the corrosion of the tested metals, as some of the wires were corroded even more using the brace cleanser. Glitter-containing test specimens showed significantly higher amounts of aluminium. Mechanical loading as well as the presence of glitter particles in the resin significantly affected ion concentrations.

[Aluminium release of glitter particles in removable orthodontic appliances]. / Freisetzung von Aluminium aus Glitzerpartikeln bei herausnehmbaren kieferorthopädischen Apparaturen.

BACKGROUND AND AIM: In order to support children's compliance with orthodontic treatment, glitter particles containing aluminium (Al) are often embedded in the acrylic of removable appliances. When worn for up to 16 h daily for 2-3 years, it can be assumed that Al ions diffuse into saliva over time. The aim of this study was to investigate the release of Al ions from the acrylic using different orthodontic wires. MATERIALS AND METHOD: Test specimens (surface area 5.65 cm2) were prepared from orthodontic resin and various wires; half contained aluminium glitter particles. The test specimens were placed in Petri dishes containing 50 ml of corrosion medium (pH 2.3) according to DIN EN ISO 10271 at 37 °C for 7 days. Inductively coupled plasma mass spectrometry (ICP-MS) was used to quantify the specific ions in the corrosion solution. RESULTS: Statistical analysis showed a significant difference in the concentration of Al ions between samples with and without glitter particles. Concentrations from samples with glitter reached up to 14,474 µg/l Al ions; samples without glitter contained on average 1260 µg/l. A small proportion of the Al ions may originate from the alloys of the wires. CONCLUSIONS: It should be investigated whether the aluminium concentration can lead to health risks for humans. In view of the findings, orthodontists should not offer appliances containing glitter in order to minimize aluminium uptake with saliva. It needs to be clarified whether the conditions found in the oral cavity lead to the same results as under the abovementioned conditions. Legislation should be developed to limit the release of aluminium from orthodontic products.

Clinical investigations of the comparability of different methods used to display occlusal contact points.

AIM: The aim of the present study was to evaluate the number, strength, and position of occlusal contacts shown using an intraoral scanner (IOS) and a digital occlusal analysis system (T-Scan) compared with the current gold standard using occlusal foil (OF). MATERIALS AND METHODS: Occlusal contacts were analyzed for 70 volunteers using OF in maximum intercuspation (MI). The contact points obtained using the IOS were evaluated using a screenshot from Zirkonzahn.Modellier CAD software. Finally, the volunteers were asked to bite on the sensor sheet of the T-Scan system. For the evaluation of these data, the contact points of the OF and the IOS were graded as light, medium, and strong. Furthermore, the positions of the contact points were analyzed for the anterior region (premolars and molars). Parametric statistical tests were applied to analyze the differences among the three methods. RESULTS: The mean number of all contact points was similar: 29 ± 8 with the OF, 30 ± 12 with the IOS, and 24 ± 10 with the T-Scan. However, results were different in terms of the grading of the strength of contact points: mean number of light contacts: 8 ± 4 OF vs 17 ± 8 IOS and 17 ± 6 T-Scan; medium contacts: 12 ± 5 OF vs 8 ± 4 IOS and 5 ± 4 T-Scan; and strong contacts: 9 ± 5 OF vs 6 ± 6 IOS and 4 ± 2 T-Scan. The positions of the occlusal contact points were also different. CONCLUSION: The data sets showed that there were differences in the distribution of occlusal contact points evaluated using the OF, the IOS, and the T-Scan system. Although the number of detected occlusal contacts was similar, different occlusal contact protocols were determined by the three different methods.

Improving d-mannitol productivity of Escherichia coli: impact of NAD, CO2 and expression of a putative sugar permease from Leuconostoc pseudomesenteroides.

The highly productive whole-cell biotransformation of D-fructose to D-mannitol with recombinant, resting cells of Escherichia coli BL21(DE3) requires the combined expression of mdh, fdh and glf which encode mannitol and formate dehydrogenases and a sugar facilitator, respectively. However, long-term stability of the system was restricted, possibly due to loss of the cofactor NAD, high concentrations of formate, formation of CO(2) affecting the internal pH of the cells, accumulation of high intracellular concentrations of D-mannitol, and export of D-mannitol. Downstream of the mdh gene of Leuconostoc pseudomesenteroides, we identified an open reading frame encoding for a putative mannitol permease. The gene was cloned and expressed in E. coli. Biochemical analyses revealed an activity as secondary carrier for D-fructose. Therefore, the carrier was named FupL and participation in D-mannitol transport was excluded. In biotransformation experiments, the productivity of D-mannitol formation obtained with the strain expressing the additional fupL gene was enhanced by 20%.

Continuous asymmetric ketone reduction processes with recombinant Escherichia coli.

The reduction of methyl acetoacetate was carried out in continuously operated biotransformation processes catalyzed by recombinant Escherichia coli cells expressing an alcohol dehydrogenase from Lactobacillus brevis. Three different cell types were applied as biocatalysts in three different cofactor regeneration approaches. Both processes with enzyme-coupled cofactor regeneration catalyzed by formate dehydrogenase or glucose dehydrogenase are characterized by a rapid deactivation of the biocatalyst. By contrast the processes with substrate-coupled cofactor regeneration by alcohol dehydrogenase catalyzed oxidation of 2-propanol could be run over a period of 7 weeks with exceedingly high substrate and cosubstrate concentrations of up to 2.5 and 2.8 mol L(-1), respectively. Even under these extreme conditions, the applied biocatalyst showed a good stability with only marginal leakage of intracellular cofactors.

Metabolic engineering of strains of Ralstonia eutropha and Pseudomonas putida for biotechnological production of 2-methylcitric acid.

In this study strains of Ralstonia eutropha H16 and Pseudomonas putida KT2440 were engineered which are suitable for biotechnological production of 2-methylcitric acid (2MC). Analysis of a previous mutant of R. eutropha able to accumulate 2MC recommended this strain as a candidate for fermentative production of 2MC. This knowledge was used for construction of strains of R. eutropha H16 and P. putida KT2440 capable of enhanced production of 2MC. In both bacteria the chromosomal genes encoding the 2-methyl-cis-aconitate hydratase (acnM) were disrupted by directed insertion of a copy of an additional 2-methylcitrate synthase gene (prpC) yielding strains R. eutropha DeltaacnM(Re)OmegaKmprpC(Pp) and P. putida DeltaacnM(Pp)OmegaKmprpC(Re). In both strains 2-methylcitrate synthase was expressed under control of the constitutive kanamycin-resistance gene (OmegaKm) resulting in up to 20-fold higher specific 2-methylcitrate synthase activities in comparison to the wild type. The disruption of the acnM gene by insertion of prpC led to a propionate- and levulinate-negative phenotype of the engineered strains, and analysis of supernatant of these strains revealed overproduction and accumulation of 2MC in the medium. A two stage cultivation regime comprising an exponential growth phase and a 2MC production phase was developed and applied to both engineered strains for optimum production of 2MC. Whereas gluconate, fructose or succinate were provided as carbon source for the exponential growth phase, a combination of propionate or levulinate as precursor substrate for provision of propionyl-CoA and succinate or fumarate as precursor substrate for provision of oxaloacetate were used in the production phase to make sure that the 2-methylcitrate synthase was provided with their substrates. Employing the optimised feeding regime P. putida DeltaacnM(Pp)OmegaKmprpC(Re) and R. eutropha DeltaacnM(Re)OmegaKmprpC(Pp) produced 2MC up to maximal concentrations of 7.2 g/L or 26.5 mM and 19.2 g/L or 70.5 mM, respectively, during 144 h of cultivation.