Visual histological grading system for the evaluation of in vitro-generated neocartilage

Here we present the development of a visual evaluation system for routine assessment of in vitro-engineered cartilaginous tissue. Neocartilage was produced by culturing human articular chondrocytes in pellet culture systems or in a scaffold-free bioreactor system. All engineered tissues were embedded in paraffin and were sectioned and stained with Safranin O-fast green. The evaluation of each sample was broken into 3 categories (uniformity and intensity of Safranin O stain, distance between cells/amount of matrix produced, and cell morphology), and each category had 4 components with a score ranging from 0 to 3. Three observers evaluated each sample, and the new system was independently tested against an objective computer-based histomorphometry system. Pellets were also assessed biochemically for glycosaminoglycan (GAG) content. Pellet histology scores correlated significantly with GAG contents and were in agreement with the computer-based histomorphometry system. This system allows a valid and rapid assessment of in vitro-generated cartilaginous tissue that has a relevant association with objective parameters indicative of cartilage quality

Stabilized high-power laser system for the gravitational wave detector advanced LIGO

An ultra-stable, high-power cw Nd:YAG laser system, developed for the ground-based gravitational wave detector Advanced LIGO (Laser Interferometer Gravitational-Wave Observatory), was comprehensively characterized. Laser power, frequency, beam pointing and beam quality were simultaneously stabilized using different active and passive schemes. The output beam, the performance of the stabilization, and the cross-coupling between different stabilization feedback control loops were characterized and found to fulfill most design requirements. The employed stabilization schemes and the achieved performance are of relevance to many high-precision optical experiments

Diffractive triangulation of radiative point sources

We describe a general method to determine the location of a point source of waves relative to a twodimensional single-crystalline active pixel detector. Based on the inherent structural sensitivity of crystalline sensor materials, characteristic detector diffraction patterns can be used to triangulate the location of a wave emitter. The principle described here can be applied to various types of waves, provided that the detector elements are suitably structured. As a prototypical practical application of the general detection principle, a digital hybrid pixel detector is used to localize a source of electrons for Kikuchi diffraction pattern measurements in the scanning electron microscope. This approach provides a promising alternative method to calibrate Kikuchi patterns for accurate measurements of microstructural crystal orientations, strains, and phase distributions

Application of a non-targeted approach by Flash Gas Chromatography-E-nose to discriminate the geographical origin of virgin olive oils

One of the fraudulent practices often applied in the olive oil sector concerns the mislabelling of the geographical origin of the products. In order to ensure that consumers are not mislead, the European Union issued a regulation concerning the definition of specific rules for the indication of geographical origin in the label (EU Reg. 29/2012). Beyond the compulsory traceability, the application of specific analytical techniques could be a useful tool to verify the conformity between the product and the information reported on the label. The aim of this work was to evaluate the effectiveness of a Flash Gas Chromatography-E-Nose, an instrument that combines functionality of electronic nose and ultra-fast GC, for the evaluation of the geographical origin of virgin olive oils (VOOs). For this purpose, more than 150 VOOs, different for their geographical origin (from single EU countries, such as Spain, Italy and Greece, and from single extra-EU countries, such as Tunisia, Turkey and Morocco) were collected and analyzed. Subsequently, a chemometric elaboration applied, with a non-targeted approach, to the chromatographic traces was realized. This permitted to build a model able to satisfactorily discriminate samples according to their geographical origin. This work is developed in the context of the project OLEUM \u201cAdvanced solutions for assuring authenticity and quality of olive oil at global scale\u201d, funded by the European Commission within the Horizon 2020 Programme (2014\u20132020, GA no. 635690). The information expressed in this abstract reflects the authors\u2019 views; the EC is not liable for the information contained therein

Stabilized lasers for advanced gravitational wave detectors

Second generation gravitational wave detectors require high power lasers with more than 100 W of output power and with very low temporal and spatial fluctuations. To achieve the demanding stability levels required, low noise techniques and adequate control actuators have to be part of the high power laser design. In addition feedback control and passive noise filtering is used to reduce the fluctuations in the so-called prestabilized laser system (PSL). In this paper, we discuss the design of a 200 W PSL which is under development for the Advanced LIGO gravitational wave detector and will present the first results. The PSL noise requirements for advanced gravitational wave detectors will be discussed in general and the stabilization scheme proposed for the Advanced LIGO PSL will be described

Molecular Markers for Genetic Diversity Studies in African Leafy Vegetables

African leafy vegetables are becoming important crops in tackling nutrition and food security in many parts of sub-Saharan Africa, since they provide important micronutrients and vitamins, and help resource-poor farm families bridge lean periods of food shortage. Genetic diversity studies are essential for crop improvement programmes as well as germplasm conservation efforts, and research on genetic diversity of these vegetables using molecular markers has been increasing over time. Diversity studies have evolved from the use of morphological and biochemical markers to molecular markers. Molecular markers provide valuable data, since they detect mostly selectively neutral variations at the DNA level. They are well established and their strengths and limitations have been described. New marker types are being developed from a combination of the strengths of the basic techniques to improve sensitivity, reproducibility, polymorphic information content, speed and cost. This review discusses the principles of some of the established molecular markers and their application to genetic diversity studies of African leafy vegetables with a main focus on the most common Solanum, Amaranthus, Cleome and Vigna species.BMBF/HORTINLEABM

Decrease in the expression of the type 1 PTH/PTHrP receptor (PTH1R) on chondrocytes in animals with osteoarthritis

<p>Abstract</p> <p>Background</p> <p>To evaluate the expression of the type 1 PTH/PTHrP receptor (PTH1R) on chondrocytes from hyaline cartilage over the course of osteoarthritis (OA).</p> <p>Methods</p> <p>In 12 NZW rabbits, the anterior cruciate ligament (ACL) was resected to create anterior instability of the knee. In 12 control rabbits, only a sham operation, without resection of the ACL, was performed. Four animals from each group were killed at 3, 6, and 12 weeks. After opening the knee joint, OA was macroscopically graded and hyaline cartilage of the load-bearing area was evaluated histologically according to the Mankin scale and by immunostaining for PTH1R.</p> <p>Results</p> <p>There was a positive linear correlation between the time after surgery and the macroscopic and histologic OA scores. The scores in the control group were constant over the time course. Immunostaining showed significantly less expression of PTH1R in the experimental compared to the control group after 6 (P < 0.05) and 12 weeks (P < 0.01). In the experimental group, a negative linear correlation between PTH1R expression and macroscopic and histologic grades was found.</p> <p>Conclusions</p> <p>The results show an in vivo decrease in the expression of PTH1R on chondrocytes over the time course of OA. Further studies are needed to evaluate whether new treatment approaches could evolve from this knowledge.</p

Genetic variants of the promoter of the heme oxygenase-1 gene and their influence on cardiovascular disease (The Ludwigshafen Risk and Cardiovascular Health Study)

Background Heme oxygenase-1 is an inducible cytoprotective enzyme which handles oxidative stress by generating anti-oxidant bilirubin and vasodilating carbon monoxide. A (GT)n dinucleotide repeat and a -413A>T single nucleotide polymorphism have been reported in the promoter region of HMOX1 to both influence the occurrence of coronary artery disease and myocardial infarction. We sought to validate these observations in persons scheduled for coronary angiography. Methods We included 3219 subjects in the current analysis, 2526 with CAD including a subgroup of CAD and MI (n = 1339) and 693 controls. Coronary status was determined by coronary angiography. Risk factors and biochemical parameters (bilirubin, iron, LDL-C, HDL-C, and triglycerides) were determined by standard procedures. The dinucleotide repeat was analysed by PCR and subsequent sizing by capillary electrophoresis, the -413A>T polymorphism by PCR and RFLP. Results In the LURIC study the allele frequency for the -413A>T polymorphism is A = 0,589 and T = 0,411. The (GT)n repeats spread between 14 and 39 repeats with 22 (19.9%) and 29 (47.1%) as the two most common alleles. We found neither an association of the genotypes or allelic frequencies with any of the biochemical parameters nor with CAD or previous MI. Conclusion Although an association of these polymorphisms with the appearance of CAD and MI have been published before, our results strongly argue against a relevant role of the (GT)n repeat or the -413A>T SNP in the HMOX1 promoter in CAD or MI

Toward a harmonized and standardized protocol for the determination of total hydroxytyrosol and tyrosol content in virgin olive oil (VOO). The pros of a fit for the purpose ultra high performance liquid chromatography (UHPLC) procedure

\u3a4oward a harmonized and standardized procedure for the determination of total hydroxytyrosol and tyrosol content in virgin olive oil (VOO), the pros of a recently published in house validated ultra high performance liquid chromatography (UHPLC) protocol are discussed comparatively with those of other procedures that determine directly or indirectly the compounds hosted under the health claim on "olive oil polyphenols" (EC regulation 432/2012). Authentic VOOs were analyzed with five different liquid chromatographic separation protocols and 1H-NMR one in five different laboratories with expertise in VOO phenol analysis within three months. Data comparison indicated differences in absolute values. Method comparison using appropriate tools (Passing-Bablok regression and Bland Altman analyses) for all protocols vs. the UHPLC one indicated slight or statistically significant differences. The results were also discussed in terms of cost effectiveness, detection means, standard requirements and ways to calculate the total hydroxytyrosol and tyrosol content. Findings point out that the in-house validated fit for the purpose UHPLC protocol presents certain pros that should be exploited by the interested parties. These are the simplicity of sample preparation, fast elution time that increase the number of samples analyzed per day and integration of well-resolved peaks with the aid of only two commercially available external standards. Importance of correction factors in the calculations is stressed