The design and validation of an optical coherence tomography-based classification system for focal vitreomacular traction

PurposeTo develop and validate a classification system for focal vitreomacular traction (VMT) with and without macular hole based on spectral domain optical coherence tomography (SD-OCT), intended to aid in decision-making and prognostication.MethodsA panel of retinal specialists convened to develop this system. A literature review followed by discussion on a wide range of cases formed the basis for the proposed classification. Key features on OCT were identified and analysed for their utility in clinical practice. A final classification was devised based on two sequential, independent validation exercises to improve interobserver variability.ResultsThis classification tool pertains to idiopathic focal VMT assessed by a horizontal line scan using SD-OCT. The system uses width (W), interface features (I), foveal shape (S), retinal pigment epithelial changes (P), elevation of vitreous attachment (E), and inner and outer retinal changes (R) to give the acronym WISPERR. Each category is scored hierarchically. Results from the second independent validation exercise indicated a high level of agreement between graders: intraclass correlation ranged from 0.84 to 0.99 for continuous variables and Fleiss' kappa values ranged from 0.76 to 0.95 for categorical variables.ConclusionsWe present an OCT-based classification system for focal VMT that allows anatomical detail to be scrutinised and scored qualitatively and quantitatively using a simple, pragmatic algorithm, which may be of value in clinical practice as well as in future research studies.

Effect of blood flow patterns on localized platelet adhesion under physiologic flow conditions using two-dimensional and three-dimensional stent models: An experimental and computational approach

This dissertation presents dynamic flow experiments with fluorescently labeled platelets to allow for spatial observation of wall attachment in inter-strut spacings, to investigate their relationship to flow patterns. Human blood with fluorescently labeled platelets was circulated through an in vitro system that produced physiologic pulsatile flow in (1) a parallel plate blow chamber that contained two-dimensional (2D) stents that feature completely recirculating flow, partially recirculating flow, and completely reattached flow, and (2) a three-dimensional (3D) cylindrical tube that contained stents of various geometric designs. ^ Flow detachment and reattachment points exhibited very low platelet deposition. Platelet deposition was very low in the recirculation regions in the 3D stents unlike the 2D stents. Deposition distal to a strut was always high in 2D and 3D stents. Spirally recirculating regions were found in 3D unlike in 2D stents, where the deposition was higher than at well-separated regions of recirculation. ^

Epidermal Growth Factor Impairs Palatal Shelf Adhesion and Fusion in the T gf-β3 Null Mutant

The cleft palate presented by transforming growth factor-β3 (Tgf-β3 ) null mutant mice is caused by altered palatal shelf adhesion, cell proliferation, epithelial-to-mesenchymal transformation and cell death. The expression of epidermal growth factor (EGF), transforming growth factor-β1 ( Tgf-β1 ) and muscle segment homeobox-1 (Msx-1) is modified in the palates of these knockout mice, and the cell proliferation defect is caused by the change in EGF expression. In this study, we aimed to determine whether this change in EGF expression has any effect on the other mechanisms altered in Tgf-β 3 knockout mouse palates. We tested the effect of inhibiting EGF activity in vitro in the knockout palates via the addition of Tyrphostin AG 1478. We also investigated possible interactions between EGF, Tgf-β 1 and Msx-1 in Tgf-β 3 null mouse palate cultures. The results show that the inhibition of EGF activity in Tgf-β 3 null mouse palate cultures improves palatal shelf adhesion and fusion, with a particular effect on cell death, and restores the normal distribution pattern of Msx-1 in the palatal esenchyme. Inhibition of TGF-β 1 does not affect either EGF or Msx-1 expression.

Fibrillar Elastomeric Micropatterns Create Tunable Adhesion Even to Rough Surfaces

Acknowledgements V.B., N.K.G., and E.A. contributed with conception and experimental design. V.B. performed the experiments. V.B., R.H., A.G., and R.M.M. carried out analysis and interpretation of data. V.B., R.H., A.G., and E.A. wrote the manuscript. V.B. and R.H. contributed equally to this work. V.B. acknowledges funding by SPP 1420 of the German Science Foundation DFG. E.A., N.K.G., and R.H. acknowledge funding from the European Research Council under the European Union/ERC Advanced Grant “Switch2Stick,” Agreement No. 340929.

Near-field diffraction-based focal length determination technique

An accurate and simple technique for determining the focal length of a lens is presented. It consists of measuring the period of the fringes produced by a diffraction grating at the near field when it is illuminated with a beam focused by the unknown lens. In paraxial approximation, the period of the fringes varies linearly with the distance. After some calculations, a simple extrapolation of data is performed to obtain the locations of the principal plane and the focal plane of the lens. Thus, the focal length is obtained as the distance between the two mentioned planes. The accuracy of the method is limited by the collimation degree of the incident beam and by the algorithm used to obtain the period of the fringes. We have checked the technique with two commercial lenses, one convergent and one divergent, with nominal focal lengths (+100±1) mm and (−100±1) mm respectively. We have experimentally obtained the focal lengths resulting into the interval given by the manufacturer but with an uncertainty of 0.1%, one order of magnitude lesser than the uncertainty given by the manufacturer.

Does the degree of coarctation of the aorta influence wall shear stress focal heterogeneity?

The development of atherosclerosis in the aorta is associated with low and oscillatory wall shear stress for normal patients. Moreover, localized differences in wall shear stress heterogeneity have been correlated with the presence of complex plaques in the descending aorta. While it is known that coarctation of the aorta can influence indices of wall shear stress, it is unclear how the degree of narrowing influences resulting patterns. We hypothesized that the degree of coarctation would have a strong influence on focal heterogeneity of wall shear stress. To test this hypothesis, we modeled the fluid dynamics in a patient-specific aorta with varied degrees of coarctation. We first validated a massively parallel computational model against experimental results for the patient geometry and then evaluated local shear stress patterns for a range of degrees of coarctation. Wall shear stress patterns at two cross sectional slices prone to develop atherosclerotic plaques were evaluated. Levels at different focal regions were compared to the conventional measure of average circumferential shear stress to enable localized quantification of coarctation-induced shear stress alteration. We find that the coarctation degree causes highly heterogeneous changes in wall shear stress.

An Infection-Responsive Approach to Reduce Bacterial Adhesion in Urinary Biomaterials

Infection is an inevitable consequence of chronic urinary catheterisation, with associated problems of recurrent catheter encrustation and blockage experienced by approximately 50% of all long-term catheterised patients. In this work we have exploited, for the first time, the reported pathogen-induced elevation of urine pH as a trigger for ‘intelligent’ antimicrobial release from novel hydrogel drug delivery systems of 2-hydroxyethyl methacrylate and vinyl-functionalised nalidixic acid derivatives, developed as candidate infection-resistant urinary catheter coatings. Demonstrating up to 20-fold faster rates of drug release at pH 10, representing infected urine pH, than at pH 7, and achieving reductions of up to 96.5% in in vitro bacterial adherence, our paradigm of pH-responsive drug delivery, which requires no external manipulation, therefore represents a promising development towards the prevention of catheter-associated urinary tract infections (CAUTIs) in vivo.

MMSET deregulation affects cell cycle progression and adhesion regulons in t(4;14) myeloma plasma cells

BackgroundThe recurrent immunoglobulin translocation, t(4;14)(p16;q32) occurs in 15% of multiple myeloma patients and is associated with poor prognosis, through an unknown mechanism. The t(4;14) up-regulates fibroblast growth factor receptor 3 (FGFR3) and multiple myeloma SET domain (MMSET) genes. The involvement of MMSET in the pathogenesis of t(4;14) multiple myeloma and the mechanism or genes deregulated by MMSET upregulation are still unclear.Design and MethodsThe expression of MMSET was analyzed using a novel antibody. The involvement of MMSET in t(4;14) myelomagenesis was assessed by small interfering RNA mediated knockdown combined with several biological assays. In addition, the differential gene expression of MMSET-induced knockdown was analyzed with expression microarrays. MMSET gene targets in primary patient material was analyzed by expression microarrays.ResultsWe found that MMSET isoforms are expressed in multiple myeloma cell lines, being exclusively up-regulated in t(4;14)-positive cells. Suppression of MMSET expression affected cell proliferation by both decreasing cell viability and cell cycle progression of cells with the t(4;14) translocation. These findings were associated with reduced expression of genes involved in the regulation of cell cycle progression (e.g. CCND2, CCNG1, BRCA1, AURKA and CHEK1), apoptosis (CASP1, CASP4 and FOXO3A) and cell adhesion (ADAM9 and DSG2). Furthermore, we identified genes involved in the latter processes that were differentially expressed in t(4;14) multiple myeloma patient samples.ConclusionsIn conclusion, dysregulation of MMSET affects the expression of several genes involved in the regulation of cell cycle progression, cell adhesion and survival.

Vasodilator-Stimulated Phosphoprotein (VASP)-dependent and -independent pathways regulate thrombin-induced activation of Rap1b in platelets

Background: Vasodilator-Stimulated Phosphoprotein (VASP) is involved in the inhibition of agonist-induced platelet aggregation by cyclic nucleotides and the adhesion of platelets to the vascular wall. αIIbβ3 is the main integrin responsible for platelet activation and Rap1b plays a key role in integrin signalling. We investigated whether VASP is involved in the regulation of Rap1b in platelets since VASP-null platelets exhibit augmented adhesion to endothelial cells in vivo.

Methods: Washed platelets from wild type and VASP-deficient mice were stimulated with thrombin, the purinergic receptors agonist ADP, or the thromboxane A2 receptor agonist U46619 and Rap1b activation was measured using the GST-RalGDS-RBD binding assay. Interaction of VASP and Crkl was investigated by co-immunoprecipitation, confocal microscopy, and pull-down assays using Crkl domains expressed as GST-fusion proteins.

Results: Surprisingly, we found that activation of Rap1b in response to thrombin, ADP, or U46619 was significantly reduced in platelets from VASP-null mice compared to platelets from wild type mice. However, inhibition of thrombin-induced activation of Rap1b by nitric oxide was similar in platelets from wild type and VASP-null mice indicating that the NO/cGMP/PKG pathway controls inhibition of Rap1b independently from VASP. To understand how VASP regulated Rap1b, we investigated association between VASP and the Crk-like protein (Crkl), an adapter protein which activates the Rap1b guanine nucleotide exchange factor C3G. We demonstrated the formation of a Crkl/VASP complex by showing that: 1) Crkl co-immunoprecipitated VASP from platelet lysates; 2) Crkl and VASP dynamically co-localized at actin-rich protrusions reminiscent of focal adhesions, filopodia, and lamellipodia upon platelet spreading on fibronectin; 3) recombinant VASP bound directly to the N-terminal SH3 domain of Crkl; 4) PKA-mediated VASP phosphorylation on Ser157 abrogated the binding of Crkl.

Conclusions: We identified Crkl as a novel protein interacting with VASP in platelets. We propose that the C3G/Crkl/VASP complex plays a role in the regulation of Rap1b and this explains, at least in part, the reduced agonist-induced activation of Rap1b in VASP-null platelets. In addition, the fact that PKA-dependent VASP phosphorylation abrogated its interaction with Crkl may provide, at least in part, a rationale for the PKA-dependent inhibition of Rap1b and platelet aggregation.

Focal length calibration for depth recovery using a focusing technique

[EN]The work presented in this paper is related to Depth Recovery from Focus The approach starts calibrating focal length of the camera using the Gaussian lens law for the thin lens camera model Two approaches are presented based on the availability of the internal distance of the lens

MicroRNA-induced polarization of microglia and macrophages after focal cerebral ischemia

Der ischämische Schlaganfall ist nicht nur die zweithäufigste Todesursache weltweit, sondern auch eine der Hauptursachen für körperliche Beeinträchtigungen im Erwachsenenalter. Das Ausmaß der durch den Schlaganfall hervorgerufenen Gewebeschädigung ist stark durch das Immunsystem geprägt. Die im Zentralnervensystem (ZNS) ansässigen Mikroglia und die aus dem Blutsystem infiltrierenden Makrophagen sind die Schlüsselzellen der lokalen und systemischen Entzündungsantwort nach dem ischämischen Schlaganfall. Sowohl Mikroglia als auch Makrophagen spielen in der Entwicklung der Gewebeschädigung eine duale Rolle. Zum einen phagozytieren sie Zelltrümmer und unterstützen neuroregenerative Prozesse, zum anderen sind diese Zellen in der Lage den Zustand der Gewebsschädigung zu verschlimmern und einer Regeneration des ZNS entgegenzuwirken. Die Polarisierung der Mikroglia/Makrophagen hin zu verschiedenen Phänotypen ist abhängig von der jeweiligen Phase der Gewebeschädigung. Der destruktive, proinflammatorische Phänotyp (M1) steht dabei dem protektiven, antiinflammatorischen Phänotyp (M2) gegenüber. Die Notwendigkeit einer zielgerichteten Regulierung der polarisierten Mikroglia/Makrophagen zum protektiven M2-Phänotyp wurde bereits mehrfach im Zusammenhang mit der Behandlung von neurodegenerativen Erkrankungen erwähnt. In der vorliegenden Dissertation soll die immunregulierende und neuroprotektive Wirkung der microRibonukleinsäure-124 (miRNA-124) in Bezug auf die Polarisierung von Mikroglia/Makrophagen zu verschiedenen Zeitpunkten nach Verschluss der Arteria cerebri media (ACM) im Gehirn von Mäusen gezeigt werden. Zu diesem Zweck wurde die liposomierte miRNA-124 zu einem frühen Zeitpunkt (Tag 2) und zu einem späten Zeitpunkt (Tag 10) nach Verschluss der ACM verabreicht. Die Behandlung mit der miRNA-124 zu einem frühen Zeitpunkt resultierte dabei in einem signifikanten Anstieg in der Anzahl der M2-positiven Mikroglia/Makrophagen im Vergleich zur Kontrollgruppe. Gleichzeitig nahm die Anzahl der M1-positiven Mikroglia/Makrophagen signifikant ab. Im Wesentlichen resultierte die Behandlung mit der miRNA-124 zu beiden Zeitpunkten in einem geringeren Verhältnis von proinflammatorischen (M1) zu antiinflammatorischen (M2) Mikroglia/Makrophagen. Zu den weiteren Erkenntnissen einer frühzeitigen Behandlung im Rahmen dieser Dissertation gehören: (i) eine signifikante Zunahme des neuronalen Überlebens, das zudem positiv mit der Anzahl der M2-positiven Mikroglia/Makrophagen korreliert, (ii) eine verbesserte funktionelle Erholung, welche in Verbindung mit den veränderten neuroinflammatorischen Ereignissen steht und (iii) ein signifikant verkleinertes Läsionsareal, welches durch reaktive Astrozyten zum gesunden Gewebe hin abgegrenzt wird. Die Ergebnisse dieser Dissertation zeigen, dass die Verabreichung von miRNA-124 eine neue Möglichkeit zur Regulierung der Immunantwort und der Neuroprotektion im Rahmen der Behandlung des ischämischen Schlaganfalls darstellt.

Inteligência coopetitiva setorial: uma proposta de modelo

The business environment context points at the necessity of new forms of management for the sustainable competitiveness of organizations through time. Coopetition is characterized as an alternative in the interaction of different actors, which compete and cooperate simultaneously, in the pursuit of common goals. This dual relation, within a gain-increasing perspective, converts competitors into partners and fosters competitiveness, especially that of organizations within a specific sector. The field of competitive intelligence has, in its turn, assisted organizations, individually, in the systematization of information valuable to decision-making processes, which benefits competitiveness. It follows that it is possible to combine coopetition and competitive intelligence in a systematized process of sectorial intelligence for coopetitive relations. The general aim of this study is, therefore, to put forth a model of sectorial coopetitive intelligence. The methodological outlining of the study is characterized as a mixed approach (quantitative and qualitative methods), of an applied nature, of exploratory and descriptive aims. The Coordination of the Strategic Roadmapping Project for the Future of Paraná's Industry is the selected object of investigation. Protocols have been designed to collect primary and secondary data. In the collection of the primary ata, online questionary were sent to the sectors selected for examination. A total of 149 answers to the online questionary were obtained, and interviews were performed with all embers of the technical team of the Coordination, in a total of five interviewees. After the collection, all the data were tabulated, analyzed and validated by means of focal groups with the same five members of the Coordination technical team, and interviews were performed with a representative of each of the four sectors selected, in a total of nine participants in the validation. The results allowed the systematization of a sectorial coopetitive intelligence model called ICoops. This model is characterized by five stages, namely, planning, collection, nalysis, project development, dissemination and evaluation. Each stage is detailed in inputs, activities and outputs. The results suggest that sectorial coopetition is motivated mainly by knowledge sharing, technological development, investment in R&D, innovation, chain integration and resource complementation. The importance of a neutral institution has been recognized as a facilitator and incentive to the approximation of organizations. Among the main difficulties are the financing of the projects, the adhesion of new members, the lack of tools for the analysis of information and the dissemination of the actions.

Advanced Adhesion Strength Testing Methods of Thin Film Multilayers in Electronic Packaging Systems

With the continued miniaturization and increasing performance of electronic devices, new technical challenges have arisen. One such issue is delamination occurring at critical interfaces inside the device. This major reliability issue can occur during the manufacturing process or during normal use of the device. Proper evaluation of the adhesion strength of critical interfaces early in the product development cycle can help reduce reliability issues and time-to-market of the product. However, conventional adhesion strength testing is inherently limited in the face of package miniaturization, which brings about further technical challenges to quantify design integrity and reliability. Although there are many different interfaces in today's advanced electronic packages, they can be generalized into two main categories: 1) rigid to rigid connections with a thin flexible polymeric layer in between, or 2) a thin film membrane on a rigid structure. Knowing that every technique has its own advantages and disadvantages, multiple testing methods must be enhanced and developed to be able to accommodate all the interfaces encountered for emerging electronic packaging technologies. For evaluating the adhesion strength of high adhesion strength interfaces in thin multilayer structures a novel adhesion test configuration called “single cantilever adhesion test (SCAT)” is proposed and implemented for an epoxy molding compound (EMC) and photo solder resist (PSR) interface. The test method is then shown to be capable of comparing and selecting the stronger of two potential EMC/PSR material sets. Additionally, a theoretical approach for establishing the applicable testing domain for a four-point bending test method was presented. For evaluating polymeric films on rigid substrates, major testing challenges are encountered for reducing testing scatter and for factoring in the potentially degrading effect of environmental conditioning on the material properties of the film. An advanced blister test with predefined area test method was developed that considers an elasto-plastic analytical solution and implemented for a conformal coating used to prevent tin whisker growth. The advanced blister testing with predefined area test method was then extended by employing a numerical method for evaluating the adhesion strength when the polymer’s film properties are unknown.

Adhesion molecules and chemokines: relation to anthropometric, body composition, biochemical and dietary variables

Introduction: Among the inflammatory mediators involved in the pathogenesis of obesity, the cell adhesion molecules P-selectin, E-selectin, VCAM-1, ICAM-1 and the chemokine MCP-1 stand out. They play a crucial role in adherence of cells to endothelial surfaces, in the integrity of the vascular wall and can be modulated by body composition and dietary pattern. Objectives: To describe and discuss the relation of these cell adhesion molecules and chemokines to anthropometric, body composition, dietary and biochemical markers. Methods: Papers were located using scientific databases by topic searches with no restriction on year of publication. Results: All molecules were associated positively with anthropometric markers, but controversial results were found for ICAM-1 and VCAM-1. Not only obesity, but visceral fat is more strongly correlated with E-selectin and MCP-1 levels. Weight loss influences the reduction in the levels of these molecules, except VCAM-1. The distribution of macronutrients, excessive consumption of saturated and trans fat and a Western dietary pattern are associated with increased levels. The opposite could be observed with supplementation of w-3 fatty acid, healthy dietary pattern, high calcium diet and high dairy intake. Regarding the biochemical parameters, they have inverse relation to HDLC and positive relation to total cholesterol, triglycerides, blood glucose, fasting insulin and insulin resistance. Conclusion: Normal anthropometric indicators, body composition, biochemical parameters and eating pattern positively modulate the subclinical inflammation that results from obesity by reducing the cell adhesion molecules and chemokines.