The Dysregulated Galectin Network Activates NF-κB to Induce Disease Markers and Matrix Degeneration in 3D Pellet Cultures of Osteoarthritic Chondrocytes.

This work aimed to study the dysregulated network of galectins in OA chondrocyte pellets, and to assess whether their recently discovered activity as molecular switches of functional biomarkers results in degradation of extracellular matrix in vitro. Scaffold-free 3D pellet cultures were established of human OA chondrocytes. Expression and secretion of galectin(Gal)-1, -3, and -8 were monitored relative to 2D cultures or clinical tissue sections by RT-qPCR, immunohistochemistry and ELISAs. Exposure of 2D and 3D cultures to an in vivo-like galectin mixture (Gal-1 and Gal-8: 5 µg/ml, Gal-3: 1 µg/ml) was followed by the assessment of pellet size, immunohistochemical matrix staining, and/or quantification of MMP-1, -3, and -13. Application of inhibitors of NF-κB activation probed into the potential of intervening with galectin-induced matrix degradation. Galectin profiling revealed maintained dysregulation of Gal-1, -3, and -8 in pellet cultures, resembling the OA situation in situ. The presence of the galectin mixture promoted marked reduction of pellet size and loss of collagen type II-rich extracellular matrix, accompanied by the upregulation of MMP-1, -3, and -13. Inhibition of p65-phosphorylation by caffeic acid phenethyl ester effectively alleviated the detrimental effects of galectins, resulting in downregulated MMP secretion, reduced matrix breakdown and augmented pellet size. This study suggests that the dysregulated galectin network in OA cartilage leads to extracellular matrix breakdown, and provides encouraging evidence of the feasible inhibition of galectin-triggered activities. OA chondrocyte pellets have the potential to serve as in vitro disease model for further studies on galectins in OA onset and progression.

Ketogenic diets in patients with inherited metabolic disorders.

Ketogenic diets (KDs) are diets that bring on a metabolic condition comparable to fasting, usually without catabolism. Since the mid-1990s such diets have been widely used in patients with seizures/epilepsies, mostly children. This review focuses on the use of KDs in patients with various inherited metabolic disorders (IMD). In glucose transporter type 1 deficiency syndrome (GLUT1-DS) and pyruvate dehydrogenase complex (PDHc) deficiency, KDs are deemed the therapy of choice and directly target the underlying metabolic disorder. Moreover, in other IMD, mainly of intermediary metabolism such as glycogen storage diseases and disorders of mitochondrial energy supply, KDs may ameliorate clinical symptoms and laboratory parameters. KDs have also been used successfully to treat symptoms such as seizures/epilepsy in IMD, e.g. in urea cycle disorders and non-ketotic hyperglycinemia. As a note of caution, catabolism may cause the condition of patients with IMD to deteriorate and should thus be avoided during KDs. For this reason, careful monitoring (clinical, laboratory and apparatus-supported) is warranted. In some IMDs specific macronutrient supply is critical. Therefore, in cases of PDHc deficiency the carbohydrate intake tolerated without lactate increase and in urea cycle disorders the protein tolerance should be determined. Considering this, it is particularly important in patients with IMD that the use of KDs be individualized and well documented.

Expression of matrix metalloproteinases in human growth plate chondrocytes is enhanced at high levels of mechanical loading: A possible explanation for overuse injuries in children.

Matrix metalloproteinases (MMPs), responsible for extracellular matrix remodelling and angiogenesis, might play a major role in the response of the growth plate to detrimental loads that lead to overuse injuries in young athletes. In order to test this hypothesis, human growth plate chondrocytes were subjected to mechanical forces equal to either physiological loads, near detrimental or detrimental loads for two hours. In addition, these cells were exposed to physiological loads for up to 24 hours. Changes in the expression of MMPs -2, -3 and -13 were investigated. We found that expression of MMPs in cultured human growth plate chondrocytes increases in a linear manner with increased duration and intensity of loading. We also showed for the first time that physiological loads have the same effect on growth plate chondrocytes over a long period of time as detrimental loads applied for a short period. These findings confirm the involvement of MMPs in overuse injuries in children. We suggest that training programmes for immature athletes should be reconsidered in order to avoid detrimental stresses and over-expression of MMPs in the growth plate, and especially to avoid physiological loads becoming detrimental.

Aquaporin 1 (AQP1) expression in experimentally induced osteoarthritic knee menisci: an in vivo and in vitro study.

Osteoarthritis (OA) of the knee is a major problem in our society. The development of new treatment options for OA is limited, because the pathophysiological mechanisms are not clearly understood, especially on the molecular level. Aquaporin 1 (AQP1) is a specific protein channels for water transport; it is expressed in articular chondrocytes, human synovitis, in chondrocytes of patients with rheumatoid arthritis or OA and in chondrocyte-like cells of human intervertebral disc. The aim of this study was to investigate the expression of AQP1, through immunohistochemistry, immunocytochemistry and Western blot, in experimentally induced OA knee menisci. AQP1 was studied in vivo in knee OA menisci from 36 rats that underwent medial or lateral meniscectomy, and in vitro on fibrochondrocytes derived from knee OA menisci rats. OA in rats was experimentally induced and tested by histomorphometric analysis. Histological results demonstrated structural alterations in OA menisci accompanied by a very strong AQP1 immunohistochemical and immunocytochemical staining. The Western blot analysis confirmed a strong expression of AQP1 in OA fibrochondrocytes cells. The results of the present research suggest that an activation of AQP1, induced by the OA process, may represent an endogenous mechanism, which can be used to control the tissue degeneration within OA articular joints.

Supraglottic combined frequency jet ventilation versus subglottic monofrequent jet ventilation in patients undergoing microlaryngeal surgery.

UNLABELLED: We compared the efficacy of gas exchange during supraglottic combined-frequency jet ventilation via a jet ventilation laryngoscope and during monofrequent jet ventilation via the Mon-Jet catheter (Xomed, Jacksonville, FL). Twenty-three anesthetized (propofol, fentanyl, vecuronium) patients undergoing microlaryngeal surgery were prospectively studied and randomly assigned to one of two groups. The patients' lungs were ventilated with combined-frequency jet ventilation (10 min, 15 and 600 breaths/min, inspiration/expiration time ratio = 1, driving pressure 750-1500 mm Hg), monofrequent (low-frequency group: 15 breaths/min; high-frequency group: 600 breaths/min) jet ventilation (20 min), and again combined-frequency jet ventilation (15 min). PaO(2), PaCO(2), and the inspiratory oxygen fraction (FIO(2)) were measured. Wilcoxon's signed rank test was applied. During monofrequent jet ventilation, PaCO(2) increased and the PaO(2)/FIO(2) decreased significantly (P < 0.05) as compared with combined-frequency jet ventilation (low-frequency group: PaCO(2) from 39.4 +/- 3.3 to 50. 8 +/- 8.0 mm Hg, PaO(2)/FIO(2) from 306 +/- 100 to 225 +/- 94 mm Hg; high-frequency group: PaCO(2) from 36.7 +/- 7.2 to 60.3 +/- 6.1 mm Hg, PaO(2)/FIO(2) from 429 +/- 87 to 190 +/- 51 mm Hg; mean +/- SD). After switching back to combined-frequency jet ventilation, PaCO(2) decreased and PaO(2)/FIO(2) increased to baseline levels. We conclude that gas exchange during microlaryngeal surgery can be more easily maintained with supraglottic combined-frequency jet ventilation than with subglottic monofrequent jet ventilation via the Mon-Jet catheter. IMPLICATIONS: This study demonstrates that the combination of high- and low-frequency supraglottic jet ventilation via a jet ventilation laryngoscope provides a better pulmonary gas exchange and allows more accurate airway pressure monitoring during microlaryngeal surgery than subglottic monofrequent jet ventilation via an endotracheal catheter.

Using human factor analysis and VR simulation techniques for the optimization of the surgical worksystem.

The introduction of minimal invasive operating techniques into the conventional surgical worksystem, requires a fundamental analysis of the resulting problems. Based on ergonomic investigations, concerning working postures, static holding work and task sequence analysis, bottlenecks of workplace design can be demonstrated. As a contribution to the optimization of workplace design, the development of a CAD-based VR simulation environment with an integrated anthropometric man-model, as a very useful tool for anthropometric system design is presented.

Evaluation of multi-allergen dipstick screening test.

In order to determine the diagnostic performance of the Quidel Allergen Screen (QAS), 43 patients characterized by standard skin prick test and intradermal testing for atopic sensitivity were evaluated, in parallel, using Phadebas RAST and QAS to detect the presence of serum IgE. The Quidel Allergen Screen and RAST were compared with the investigator's clinical assessment. The differences between QAS and RAST are not statistically significant. The QAS appears to be equal to the Phadebas RAST in sensitivity and specificity and thus offers a rapid, easily interpreted in vitro method for the detection of specific serum IgE.