Low molecular weight polysialic acid prevents lipopolysaccharide-induced inflammatory dopaminergic neurodegeneration in humanized SIGLEC11 transgenic mice.

Parkinson's disease is one of the most common neurodegenerative diseases in the elderly population, with a pathophysiology linked to neuroinflammation, complement activation, and oxidative damage. Soluble polysialic acid with an average degree of polymerization 20 (polySia avDP20) prevents inflammation and oxidative burst in human macrophages via sialic acid-binding immunoglobulin like lectin-11 (SIGLEC11) receptor and interferes with alternative complement activation. Here, we confirmed the anti-inflammatory capacity of polySia avDP20 on cultured murine embryonic stem cell-derived microglia and analyzed the effect of polySia avDP20 in a lipopolysaccharide-triggered animal model of Parkinson's disease. We demonstrated a neuroprotective effect of intraperitoneally applied polySia avDP20 in humanized SIGLEC11 transgenic mice after repeated systemic challenge with lipopolysaccharide. Pathway enrichment analysis of the brain transcriptome on day 19 after disease initiation showed that intraperitoneal application of 10 µg/g body weight polySia avDP20 prevented excessive inflammation. In line with these data, polySia avDP20 attenuated the lipopolysaccharide-triggered increase in mRNA levels of immune-related genes (Il1b, Cd14, Myd88, Fcer1g, Itgam, C4, Cybb, Iba1 and Cd68) and cell death-related genes (Casp8, Ripk1 and Ripk3) in the brains of SIGLEC11 transgenic mice on day 19, but not on day 5. Moreover, immunohistochemistry demonstrated that polySia avDP20 reduced the lipopolysaccharide-induced increase in immunoreactivity of IBA1 and CD68 in the substantia nigra pars reticulata in SIGLEC11 transgenic and wild type mice on day 19. Furthermore, treatment with polySia avDP20 prevented the loss of dopaminergic neurons in the substantia nigra pars compacta induced by lipopolysaccharide challenge in both SIGLEC11 transgenic and wild type mice on day 19. Thus, our data demonstrate that polySia avDP20 ameliorates inflammatory dopaminergic neurodegeneration and therefore is a promising drug candidate to prevent Parkinson's disease-related inflammation and neurodegeneration.

Late relapse in patients with diffuse large B-cell lymphoma: impact of rituximab on their incidence and outcome.

Diffuse large B-cell lymphoma (DLBCL) constitutes 25-35% of all non-Hodgkin lymphomas in Western countries. Approximately two thirds of the patients can be cured with standard immuno-chemotherapy. Most relapses occur within 1-2 years from diagnosis, however, the occurrence of relapses after 5 years or more has been described. We aimed at defining the incidence and clinical features of late relapses. Data of 1113 DLBCL patients were analysed. Among the 196 patients relapsing after a first complete remission, 36 (18% of relapses and 3% of all DLBCLs) experienced a recurrence more than 5 years from diagnosis. Late relapsing patients, in comparison with those relapsing earlier, showed a more favourable risk profile at presentation: normal lactate dehydrogenase levels (P = 0·002), early Ann Arbor stage (P = 0·006) and low International Prognostic Index (P = 0·003). The risk of late relapse was lowered by the introduction of rituximab as part of the front-line treatment (P < 0·001). Cause-specific survival (CSS) from the time of relapse was significantly better for late relapsing patients compared to those relapsing early: 5-year CSS rates were 53% and 31%, respectively (P = 0·033). A trend toward a better overall survival was also observed, with 5-year rates after relapse of 47% and 25%, respectively (P = 0·054).

Apical root resorption due to mandibular first molar mesialization: A split-mouth study.

INTRODUCTION: Our aim was to evaluate the risk of external apical root resorption (EARR) in mesialized mandibular molars due to space closure in patients with unilateral second premolar agenesis. The contralateral side served as the control. METHODS: After application of eligibility criteria, 25 retrospectively selected subjects (median age, 14.9 years; range, 12.0-31.9 years) were analyzed. Space closure (approximately 10 mm) was performed using skeletal anchorage. EARR was measured at the mandibular permanent canines, first premolars, and first molars in the pretreatment and posttreatment orthopantomograms. Measurements were performed by 2 examiners independently and were corrected for distortion and magnification of radiographs, which were assessed in a pilot study. Multivariate analysis of covariance and pairwise comparisons were performed. RESULTS: The mean enlargement factor of the panoramic machine was 29% ± 0.3%. Distortion exceeded 5% only in cases of large positioning errors (>20°). Intraclass correlation coefficients showed strong to almost perfect agreement (mean, 0.80 mm; 95% CI, 0.75-0.85) of the two examiners. Multivariate analysis of covariance resulted in no difference in EARR between the canines and premolars of the space closure and control sides. On the contrary, there was a statistically significant difference between mesialized and nonmezialized molars (0.73 mm; 95% confidence interval, 0.19-1.27). The mean total EARR in each tooth type did not exceed 1 mm. CONCLUSIONS: Space closure through extensive tooth movement in the mandible was identified as a risk factor for EARR. However, the amount of EARR attributed to space closure and the total EARR were not considered clinically significant.

Only EBUS-Guided Mediastinal Lymph Node Cryobiopsy Enabled Immunotherapy in a Patient with Non-Small Cell Lung Cancer.

Personalized treatment of metastatic non-squamous non-small cell lung cancer (NSCLC) requires detailed molecular characterization of the tumour including detection of predictive driver mutations and programmed death ligand 1 (PD-L1) expression. Complete detection is influenced by the amount of tumour cells sampled as well as their quality. Different sampling techniques may be necessary to provide sufficient tumour material for comprehensive molecular characterization. Missing the detection of targetable molecular genetic aberrations would have a serious impact on the quality of life and prognosis of a patient. This case report highlights the importance of biopsy technique in a patient with NSCLC. Several procedures-pleural puncture, transthoracic lung biopsy and endobronchial ultrasound-guided transbronchial needle aspiration (EBUS-TBNA)-could not provide sufficient tumour material for precise tumour characterization. Only the addition of EBUS-guided transbronchial lymph node cryobiopsy (EBUS-TBLNC) enabled complete immunohistochemical and genetic tumour characterization, demonstrating PD-L1 expression in 100% of the tumour cells in the absence of actionable genetic alterations. Based on these results, immunotherapy was initiated.

Intraoral Scanners for In Vivo 3D Imaging of the Gingiva and the Alveolar Process.

This study aimed to assess the reliability of two intraoral surface scanners for the representation of the alveolar process in vivo. Complete maxillary scans (CS 3600, Carestream and TRIOS 3, 3Shape) were repeatedly obtained from 13 fully dentate individuals. Scanner precision and agreement were tested using 3D surface superimpositions on the following reference areas: the buccal front teeth area, the entire dental arch, the entire alveolar process, or single teeth by applying an iterative closest point algorithm. Following each superimposition, the mean absolute distance (MAD) between predefined 3D model surfaces was calculated. Outcomes were analyzed through non-parametric statistics and the visualization of color-coded distance maps. When superimpositions were performed on the alveolar process, the median scanner precision was below 0.05 mm, with statistically significant but negligible differences between scanners. The agreement between the scanners was approximately 0.06 mm. When single-tooth superimpositions were used to assess the precision of adjacent alveolar soft-tissue surfaces, the median error was 0.028 mm, and there was higher agreement between the scanners. The in vivo reliability of the intraoral scanners in the alveolar surface area was high overall. Single-tooth superimpositions should be preferred for the optimal assessment of neighboring alveolar surface areas relative to the dentition.

Low molecular weight polysialic acid binds to properdin and reduces the activity of the alternative complement pathway.

Sialic acids as the terminal caps of the cellular glycocalyx play an essential role in self-recognition and were shown to modulate complement processes via interaction between α2,3-linked sialic acids and complement factor H. Previously, it was suggested that low molecular weight α2,8-linked polysialic acid (polySia avDP20) interferes with complement activation, but the exact molecular mechanism is still unclear. Here, we show that soluble polySia avDP20 (molecular weight of ~ 6 kDa) reduced the binding of serum-derived alternative pathway complement activator properdin to the cell surface of lesioned Hepa-1c1c7 and PC-12 neuroblastoma cells. Furthermore, polySia avDP20 added to human serum blocked the alternative complement pathway triggered by plate-bound lipopolysaccharides. Interestingly, no inhibitory effect was observed with monosialic acid or oligosialic acid with a chain length of DP3 and DP5. In addition, polySia avDP20 directly bound properdin, but not complement factor H. These data show that soluble polySia avDP20 binds properdin and reduces the alternative complement pathway activity. Results strengthen the previously described concept of self-recognition of sialylation as check-point control of complement activation in innate immunity.

Intraoral scanners for capturing the palate and its relation to the dentition.

Proper superimposition of intraoral scan generated 3D models enables detailed assessment of soft and hard tissue surface changes. This requires accurate 3D models and stable structures as superimposition references. In the maxilla, different reference areas have been proposed, mostly located at the palatal region. In this in vivo study we evaluated the precision of two intraoral scanners (TRIOS 3, 3Shape and CS 3600, Carestream) at the maxilla, focusing on the palate itself and also on its spatial relation to the dentition, following palatal superimposition. Precision was tested through the superimposition of repeated scans on the palate and the dental arch. Overall, the median precision of both scanners was high (< 0.1 mm). Scanner precision was comparable when the palatal area was tested individually. However, TRIOS 3 showed higher precision regarding the assessment of the dental arch, following superimposition of repeated models on the palate (median difference: approximately 40 µm). In few cases, local areas of higher imprecision were present for both scanners, exceeding 0.3 mm. Thus, scanner precision seems to be high in small, but slightly reduced considering larger areas, with differences between scanners. However, the effect on individual tooth position relative to the palate was for both scanners limited.

Trueness and precision of intraoral scanners in the maxillary dental arch: an in vivo analysis.

Intraoral three-dimensional imaging has gained great interest in dentistry as a mean to generate risk-free imprints of the oral cavity. Accurate intraoral models facilitate proper diagnosis, growth assessment, outcome evaluation, and 3D printing applications. Here, in an actual clinical setup on 12 subjects, we evaluate the trueness and precision of two widely used intraoral scanners (TRIOS 3, 3Shape and CS 3600, Carestream), using an industrial scanner (Artec Space Spider) as a reference. Surface based matching was implemented using the iterative closest point algorithm (ICP). Trueness of the intraoral scans was analyzed by measuring their distance from the reference scan, in the upper buccal front area. Precision was tested through the distance of repeated scans regarding the whole dental arch, following superimpositions in the buccal front and in the whole dental arch area. TRIOS 3 displayed slightly higher precision (approximately 10 µm) compared to CS 3600, only after superimposition on the whole dental arch (p < 0.05). Both intraoral scanners showed good performance and comparable trueness (median: 0.0154 mm; p> 0.05). However, in individual cases and in various, not spatially defined areas, higher imprecision was evident. Thus, the intraoral scanners' appropriateness for highly demanding, spatially extended clinical applications remains questionable.

A computational approach for the identification of small GTPases based on preprocessed amino acid sequences.

The prediction of essential biological features based on a given protein sequence is a challenging task in computational biology. To limit the amount of in vitro verification, the prediction of essential biological activities gives the opportunity to detect so far unknown sequences with similar properties. Besides the application within the identification of proteins being involved in tumorigenesis, other functional classes of proteins can be predicted. The prediction accuracy depends on the selected machine learning approach and even more on the composition of the descriptor set used. A computational approach based on feedforward neural networks was applied for the prediction of small GTPases. Consequently, this was realized by taking secondary structure and hydrophobicity information as a preprocessing architecture and thus, as descriptors for the neural networks. We developed a neural network cluster, which consists of a filter network and four subfamily networks. The filter network was trained to identify small GTPases and the subfamily networks were trained to assign a small GTPase to one of the subfamilies. The accuracy of the prediction, whether a given sequence represents a small GTPase is very high (98.25%). The classifications of the subfamily networks yield comparable accuracy. The high prediction accuracy of the neural network cluster developed, gives the opportunity to suggest the use of hydrophobicity and secondary structure prediction in combination with a neural network cluster, as a promising method for the prediction of essential biological activities.

Effect of mandibular first molar mesialization on alveolar bone height: a split mouth study.

OBJECTIVES: To evaluate the risk of vertical alveolar bone loss (ABL) in mesialized mandibular permanent molars due to space closure in patients with unilateral second premolar agenesis. The contralateral side served as control. SUBJECTS AND METHODS: Twenty-five retrospectively selected subjects (median age 14.9, range 12.0, 31.9 years) were analyzed. Space closure (approximately 10 mm) was performed using skeletal anchorage. ABL was measured at mesial and distal sites of first molars in pre- and post-treatment panoramic radiographs. Measurements were corrected for distortion and magnification of radiographs. Molar angulation according to the occlusal plane was also evaluated. Permutational multivariate analysis of covariance (MANCOVA), followed by pairwise comparisons, was performed. RESULTS: MANCOVA resulted in no difference in ABL between the distal sites of mesialized molars and the control sites. On the contrary, there was statistically higher ABL, at the mesial sites of mesialized versus non-mesialized molars (p = 0.042; median 0.19 mm; range - 0.82, 1.33); though the difference was not clinically relevant. In the space closure side, mesially, only two patients had ABL higher that 1 mm. No patient had a severe bone level height defect (> 3 mm distance from the cementoenamel junction) at any point. When testing differences in molar angulation between sites and from pre- to post-treatment condition, no significant difference was detected (p > 0.05, median - 1.9°, range - 13.5, 6.2). LIMITATIONS: This is a retrospective study on panoramic radiographs. CONCLUSIONS: Space closure through extensive tooth movement was identified as a risk factor for vertical ABL, at the mesial sites of mandibular first molars. However, the amount of ABL was not clinically relevant, and thus this treatment option is considered safe in terms of ABL.

Charged aerosol detector HPLC as a characterization and quantification application of biopharmaceutically relevant polysialic acid from E. coli K1.

Polysialic acid (polySia) is widely investigated in various biopharmaceutical applications (e.g. treatment of inflammatory neurodegenerative diseases), whereby a certain polySia chain length with an average degree of polymerization 20 (polySia avDP20) shows most promising effects. In this study, a rapid analytical method using a HPLC and charged aerosol detector (CAD) for the direct chain length characterization of biopharmaceutically relevant polySia was developed. It was evaluated as a fast alternative to the commonly used 1,2-diamino-4,5-methylenedioxybenzene (DMB) HPLC application. In contrast to HPLC-FLD, the CAD-application provides the actual chain length of polySia within ∼3 h. The reliability of the HPLC-CAD was evaluated with a commercial reference sample of known chain length and biotechnologically produced LC polySia (long chain polySia with a DP ∼130). Moreover, HPLC-CAD was successfully applied in the direct detection of oligo- and polySia until DP ∼65 and can be used to monitor the thermal hydrolysis and subsequent chromatographic isolation of polySia avDP20 (average degree of polymerization 20) without DMB sample derivatization. In addition, CAD was successfully applied for polySia quantification using a modified elution gradient. It was tested as a fast alternative to commonly used thiobarbituric acid (TBA) assay. A differentiation between LC polySia and smaller, hydrolysed polySia chains was intended and possible. For LC polySia and polySia avDP20, a quadratic relation between polySia mass-concentration and CAD signal was observed. In case of LC polySia, a quadratic dependency with a determination coefficient of R2 = 0.99 in a broad concentration range between 0.025 and 15 mg mL-1 was determined. Quantification of polySia avDP20 was found to have quadratic dependency with a determination coefficient of R2 = 0.99 in a concentration range between 0.02 and 0.25 mg mL-1. The HPLC-CAD was tested for quantification with polySia references of known concentration and showed high accordance with a concentration deviation ≤6.7%. The CAD quantification method was also applied in the polySia avDP20 production process and was compared to the TBA assay. Results of a correlation plot showed a high determination coefficient of R2 = 0.98. Overall, HPLC-CAD analysis was successfully tested as a suitable characterization and quantification application in the biopharmaceutical production of polySia.

ISL1 is a major susceptibility gene for classic bladder exstrophy and a regulator of urinary tract development.

Previously genome-wide association methods in patients with classic bladder exstrophy (CBE) found association with ISL1, a master control gene expressed in pericloacal mesenchyme. This study sought to further explore the genetics in a larger set of patients following-up on the most promising genomic regions previously reported. Genotypes of 12 markers obtained from 268 CBE patients of Australian, British, German Italian, Spanish and Swedish origin and 1,354 ethnically matched controls and from 92 CBE case-parent trios from North America were analysed. Only marker rs6874700 at the ISL1 locus showed association (p = 2.22 × 10-08). A meta-analysis of rs6874700 of our previous and present study showed a p value of 9.2 × 10-19. Developmental biology models were used to clarify the location of ISL1 activity in the forming urinary tract. Genetic lineage analysis of Isl1-expressing cells by the lineage tracer mouse model showed Isl1-expressing cells in the urinary tract of mouse embryos at E10.5 and distributed in the bladder at E15.5. Expression of isl1 in zebrafish larvae staged 48 hpf was detected in a small region of the developing pronephros. Our study supports ISL1 as a major susceptibility gene for CBE and as a regulator of urinary tract development.

Computational Design of a DNA- and Fc-Binding Fusion Protein.

Computational design of novel proteins with well-defined functions is an ongoing topic in computational biology. In this work, we generated and optimized a new synthetic fusion protein using an evolutionary approach. The optimization was guided by directed evolution based on hydrophobicity scores, molecular weight, and secondary structure predictions. Several methods were used to refine the models built from the resulting sequences. We have successfully combined two unrelated naturally occurring binding sites, the immunoglobin Fc-binding site of the Z domain and the DNA-binding motif of MyoD bHLH, into a novel stable protein.